Appendix 3

Claim 1
A chemically-inducible plant gene expression cassette comprising:

a first promoter operatively linked to the alcR regulator sequence obtainable from Aspergillus nidulans which encodes the AlcR regulator protein, and
an inducible promoter operatively linked to a target gene,
the inducible promoter being activated by the regulator protein in the presence of an effective exogenous inducer whereby application of the inducer causes expression of the target gene.

Claim 11
A method for controlling plant gene expression comprising:

transforming a plant cell with a chemically-inducible plant gene expression cassette which has
a first promoter operatively linked to the alcR regulator sequence obtainable from Aspergillus nidulans which encodes the AlcR regulator protein, and
an inducible promoter operatively linked to a target gene,
the inducible promoter being activated by the regulator protein in the presence of an effective exogenous inducer whereby application of the inducer causes expression of the target gene.

Claim 12
A chimeric promoter comprising an upstream region containing

a promoter regulatory sequence obtainable from the alcA gene promoter of Aspergillus nidulans and
a downstream region containing a transcription initiation sequence, characterized in that
said upstream and downstream regions are heterologous,
the promoter is chemically-inducible and
the transcription initiation sequence is obtainable from the core promoter region of a promoter which is active in plant cells.

Claim 1
A polynucleotide comprising a nucleotide sequence encoding a tetracycline transactivator fusion protein,
said protein comprising a prokaryotic tet repressor and a eucaryotic transcriptional activator protein, and
said polynucleotide molecule being operably linked to an inducible minimal promoter,
which promoter contains at least one tet operator sequence.

Claim 23
A method to inhibit expression of a heterologous protein in a eucaryotic cell comprising:

a) obtaining a eucaryotic cell comprising:

1. a first polynucleotide molecule encoding a tetracycline transactivator fusion protein,
   said protein comprising a procaryotic tet repressor and a eucaryotic transcriptional activator protein, and
   said polynucleotide molecule being operably linked to an inducible minimal promoter, which promoter contains at least one tet operator sequence;
2. a second polynucleotide molecule encoding the heterologous protein,
   said second polynucleotide molecule being operably linked to an inducible minimal promoter, and
   said promoter containing at least one tet operator sequence; and

b) cultivating the eucaryotic cell in a medium comprising tetracycline or a tetracycline analogue such that expression of the heterologous protein is inhibited.

Claim 25
A method to enhance the expression of a heterologous protein in a eucaryotic cell comprising:

a) obtaining a eucaryotic cell comprising:

1. a first polynucleotide molecule encoding a tetracycline transactivator fusion protein,
   said protein comprising a prokaryotic tet repressor and a eucaryotic transcriptional activator protein, and
   said polynucleotide molecule being operably linked to an inducible minimal promoter, which promoter contains at least one tet operator sequence;
2. a second polynucleotide molecule encoding the heterologous protein,
   said second polynucleotide molecule being operably linked to an inducible minimal promoter, and
   said promoter containing at least one tet operator sequence; and

b) cultivating the eucaryotic cell in a medium lacking tetracycline or a tetracycline analogue such that expression of the heterologous protein is enhanced.

Claim 26
A method to activate the expression of a heterologous protein in a eucaryotic cell comprising:

a) obtaining a eucaryotic cell comprising:

1. a first polynucleotide molecule encoding a tetracycline transactivator fusion protein,
   said protein comprising a prokaryotic tet repressor and a eucaryotic transcriptional activator protein, and
   said polynucleotide molecule being operably linked to an inducible minimal promoter, which promoter contains at least one tet operator sequence;
2. a second polynucleotide molecule encoding the heterologous protein,
   said second polynucleotide molecule being operably linked to an inducible minimal promoter, and
   said promoter containing at least one tet operator sequence; and

b) cultivating the eucaryotic cell in a medium lacking tetracycline or a tetracycline analogue such that expression of the heterologous protein is activated.

Claim 27
A kit comprising a carrier means having in close confinement therein at least two container means, wherein

a) a first container means contains a first polynucleotide molecule encoding a tetracycline transactivator fusion protein,
said protein comprising a procaryotic tet repressor and a eucaryotic transcriptional activator protein, and
said polynucleotide molecule being operably linked to an inducible minimal promoter, which promoter contains at least one tet operator sequence; and
b) a second container means contains a second polynucleotide molecule encoding said inducible minimal promoter,
which promoter contains at least one tet operator sequence,
which tet operator sequence is strategically positioned for being operably linked to a heterologous polynucleotide sequence encoding a polypeptide.

Claim 28
A kit comprising a carrier means having in close confinement therein at least two container means, wherein

a) a first container means contains a eucaryotic cell transfected with a first polynucleotide molecule encoding a tetracycline transactivator fusion protein,
said protein comprising a procaryotic tet repressor and a eucaryotic transcriptional activator protein, and
said polynucleotide molecule being operably linked to an inducible minimal promoter, which promoter contains at least one tet operator sequence; and
b) a second container means contains a second polynucleotide molecule comprising an inducible minimal promoter,
which promoter contains at least one tet operator sequence,
which tet operator sequence is strategically positioned for being operably linked to a heterologous polynucleotide sequence encoding a heterologous pol ypeptide.

Claim 1
A composition of matter comprising a polynucleotide molecule encoding a tetracycline transactivator fusion protein,
said protein comprising a prokaryotic tet repressor and a eucaryotic transcriptional activator protein, and
said polynucleotide molecule being operably linked to an inducible minimal promoter, which promoter contains at least one tet operator sequence.

Claim 11
A method to decrease or shut off expression of a heterologous protein comprising:

a) transforming a eucaryotic cell with

1. a first polynucleotide molecule encoding a tetracycline transactivator fusion protein,
   said protein comprising a prokaryotic tet repressor and a eucaryotic transcriptional activator protein, and
   said polynucleotide molecule being operably linked to an inducible minimal promoter, which promoter contains at least one tet operator sequence;
2. a second polynucleotide molecule encoding the heterologous protein,
   said protein being operably linked to an inducible minimal promoter, and
   said promoter containing at least one tet operator sequence; and

b) cultivating the eucaryotic cell in a medium comprising tetracycline or a tetracycline analogue.

Claim 13
A method to activate or enhance the expression of a heterologous protein comprising:

a) transforming a eucaryotic cell with

i. a first polynucleotide molecule encoding tetracycline transactivator fusion protein,
said protein comprising a prokaryotic tet repressor and a eucaryotic transcriptional activator protein, and
said polynucleotide molecule being operably linked to an inducible promoter, which promoter contains at least one tet operator sequence;

ii. a second polynucleotide molecule encoding the heterologous protein,
said protein being operably linked to an inducible minimal promoter, and
said promoter containing at least one tet operator sequence; and

b) cultivating the eucaryotic cell in a medium lacking tetracycline or a tetracycline analogue.

Claim 17
A composition of matter consisting essentially of the plasmid pTet-Splice.

Claim 18
A composition of matter consisting essentially of the plasmid pTet-tTAK.

Claim 19
A kit comprising a carrier means having in close confinement therein at least two container means, wherein

a) a first container means contains a first polynucleotide molecule encoding a tetracycline transactivator fusion protein,
said protein comprising a procaryotic tet repressor and a eucaryotic transcriptional activator protein, and
said polynucleotide molecule being operably linked to an inducible minimal promoter, which promoter contains at least one tet operator sequence; and
b) a second container means contains a second polynucleotide molecule encoding said inducible minimal promoter,
which promoter contains at least one tet operator sequence, which tet operator sequence is strategically positioned for being operably linked to a heterologous polynucleotide sequence encoding a polypeptide.

Claim 20
A kit comprising a carrier means having in close confinement therein at least two container means, wherein

a) a first container means contains a eucaryotic cell transfected with a first polynucleotide molecule encoding a tetracycline transactivator fusion protein,
said protein comprising a procaryotic tet repressor and a eucaryotic transcriptional activator protein, and
said polynucleotide molecule being operably linked to an inducible minimal promoter, which promoter contains at least one tet operator sequence; and
b) a second container means contains a second polynucleotide molecule comprising an inducible minimal promoter,
which promoter contains at least one tet operator sequence, which tet operator sequence is strategically positioned for being operably linked to a heterologous polynucleotide sequence encoding a heterologous polypeptide.

Claim 1
A polynucleotide molecule coding for a transactivator fusion protein comprising:

a) a prokaryotic Tet repressor and
b) a eucaryotic transcriptional activator protein domain.

Claim 10
A polynucleotide molecule coding for a protein, wherein said polynucleotide is operably linked to a minimal promoter and at least one tet operator sequence.

Claim 16
A eucaryotic cell transfected with

a) a first polynucleotide molecule coding for a transactivator fusion pr otein comprising:
(i) a prokaryotic Tet repressor and
(ii) a eucaryotic transcriptional activator protein domain; and
b) a second polynucleotide molecule coding for a protein, wherein said second polynucleotide molecule is operably linked to a minimal promoter and at least one tet operator sequence.

Claim 18
A kit comprising a carrier means having in close confinement therein at least two container means, wherein

a) a first container means contains a first polynucleotide molecule coding for a transactivator fusion protein comprising:
(i) a prokaryotic Tet repressor and
(ii) a eucaryotic transcriptional activator protein domain, and
b) a second container means contains a second polynucleotide molecule comprising a minimal promoter operably linked to at least one tet operator sequence, wherein said minimal promoter is capable of being ligated to a heterologous gene sequence coding for a polypeptide.

Claim 19
A kit comprising a carrier means having in close confinement therein at least two container means, wherein

a) a first container means contains a eucaryotic cell transfected with a first polynucleotide molecule coding for a transactivator fusion protein comprising:
(i) a prokaryotic Tet repressor and
(ii) a eucaryotic transcriptional activator protein domain, and
b) a second container means contains a second polynucleotide molecule comprising a minimal promoter operably linked to at least one tet operator sequence, wherein said minimal promoter is capable of being ligated to a heterologous gene sequence coding for a polypeptide.

Claim 1
An isolated DNA molecule for integrating a polynucleotide sequence encoding a tetracycline-controllable transactivator (tTA) at a predetermined location in a second target DNA molecule,
the tTA comprising a prokaryotic Tet repressor operably linked to a polypeptide which directly or indirectly activates transcription in eucaryotic cells,
the DNA molecule comprising a polynucleotide sequence encoding the tTA flanked at 5' and 3' ends by additional polynucleotide sequences of sufficient length for homologous recombination between the DNA molecule and the second target DNA molecule at a predetermined location.

Claim 11
An isolated DNA molecule for integrating a polynucleotide sequence encoding a tetracycline-controllable transactivator (tTA) and a tTA-responsive promoter within a predetermined gene of interest in a second target DNA molecule, the DNA molecule comprising:

a) a first polynucleotide sequence comprising a 5' flanking regulatory region of the gene of interest, operably linked to;
b) a second polynucleotide sequence encoding a tTA,
the tTA comprising a prokaryotic Tet repressor operably linked to a polypeptide which directly or indirectly activates transcription in eucaryotic cells; and
c) a third polynucleotide sequence comprising a tTA-responsive promoter, operably linked to;
d) a fourth polynucleotide sequence comprising at least a portion of a coding region of the gene of interest;
wherein the first and fourth polynucleotide sequences are of sufficient length for homologous recombination between the DNA molecule and the gene of interest in the second target DNA molecule such that expression of the tTA is controlled by 5' regulatory elements of the gene of interest and expression of the gene of interest is controlled by the tTA-responsive promoter.

Claim 1
Worded the same as claim 1 of US 5 650 298.

Claim 11
Worded the same as claim 11 of US 5 650 298.

Claim 42
Use of tetracycline or a tetracycline analogue for the inhibition of a second transgene in a transgenic animal, said animal having:

a) a first transgene comprising a polynucleotide sequence encoding a tetracycline-controllable transactivator (tTA),
the tTA comprising a prokaryotic Tet repressor operably linked to a polypeptide which directly or indirectly activates transcription in eucaryotic cells; and
b) the second transgene comprising a gene of interest operably linked to a tTA-responsive promoter.

Claim 43
The use of tetracycline or a tetracycline analogue for the inhibition of transcription of a second transgene in a transgenic animal, said animal having:

a) a polynucleotide sequence encoding a tetracycline-controllable transactivator (tTA),
the tTA comprising a prokaryotic Tet repressor operably linked to a polypeptide which directly or indirectly activates transcription in eucaryotic cells, wherein the first transgene is integrated by homologous recombination at a predetermined location within a chromosome within cells of the animal; and
b) the second transgene comprising a gene of interest operably linked to a tTA-responsive promoter.

Claim 44
Use of tetracycline or a tetracycline analogue for inhibiting transcription of the gene of interest in a transgenic animal having a transgene comprising a polynucleotide sequence encoding a tetracycline-controllable transactivator (tTA) and a tTA-responsive promoter,
wherein the transgene is integrated by homologous recombination at a predetermined location within a gene of interest within cells of the animal such that expression of the tTA is controlled by 5' regulatory elements of the gene of interest and expression of the gene of interest is controlled by the tTA-responsive promoter.

Claim 48
A method for producing a non-human transgenic animal comprising:

a) introducing a DNA molecule encoding the tTA into a fertilized oocyte;
b) implanting the fertilized oocyte in a pseudopregnant foster mother; and
c) allowing the fertilized oocyte to develop into the non-human transgenic animal to thereby produce the non-human transgenic animal.

Claim 1
Worded the same as Claim 1 of Canadian patent and US 5 650 298.

Claim 11
Worded the same as Claim 11 of Canadian pat ent and US 5 650 298.

Claim 42
A non-human transgenic animal having a transgene comprising
a polynucleotide sequence encoding a tetracycline-controllable transactivator (tTA),
the tTA comprising a prokaryotic Tet repressor operably linked to a polypeptide which directly or indirectly activates transcription in eucaryotic cells.

Claim 52
A non-human transgenic animal having a transgene comprising
a polynucleotide sequence encoding a tetracycline-controllable transactivator (tTA),
the tTA comprising a prokaryotic Tet repressor operably linked to a polypeptide which directly or indirectly activates transcription in eucaryotic cells,
wherein the transgene is integrated by homologous recombination at a predetermined location within a chromosome within cells of the animal.

Claim 55
A transgenic animal having a transgene comprising
a polynucleotide sequence encoding a tetracycline-controllable transactivator (tTA) and a tTA-responsive promoter,
wherein the transgene is integrated by homologous recombination at a predetermined location within a gene of interest within cells of the animal such that expression of the tTA is controlled by 5' regulatory elements of the gene of interest and expression of the gene of interest is controlled by the tTA-responsive promoter.

Claim 1
An isolated nucleic acid molecule encoding a fusion protein which regulates transcription,
the fusion protein comprising a Tet repressor having at least one amino acid mutation that confers on the fusion protein an ability to bind a class B tet operator sequence having a nucleotide substitution at position +4 or +6, operatively linked to a polypeptide which regulates transcription in eukaryotic cells.

Claim 13
A method for regulating transcription of a tet operator-linked gene in an isolated cell, comprising:

a) introducing into the isolated cell a nucleic acid molecule encoding a fusion protein which regulates transcription,
the fusion protein comprising a Tet repressor having at least one amino acid mutation that confers on the fusion protein an ability to bind a class B tet operator sequence having a nucleotide substitution at position 14 or 16, operatively linked to a polypeptide which regulates transcription in eukaryotic cells; and
b) modulating the concentration of a tetracycline, or analogue thereof, in contact with the isolated cell.

Claim 1
An isolated nucleic acid encoding a fusion protein which activates transcription, the fusion protein comprising
a first polypeptide which binds to a tet operator sequence in the presence of tetracycline or a tetracycline analogue operatively linked
to a second polypeptide which activates transcription in eukaryotic cells.

Claim 26
A kit comprising a carrier means having in close confinement therein at least two container means comprising:

a) a first container means containing a first nucleic acid encoding a fusion protein which activates transcription, the fusion protein comprising
a polypeptide which binds to a first class of tet operator sequence in the presence of tetracycline or a tetracycline analogue operatively linked to a polypeptide which activates transcription in eukaryotic cells; and
b) a second container means containing a second nucleic acid comprising a first cloning site for introduction of a first nucleotide sequence to be transcribed operatively linked to at least one tet operator sequence of a first class type.

Claim 1
An isolated nucleic acid encoding a fusion protein which inhibits transcription in eukaryotic cells, the fusion protein comprising a first polypeptide which binds to tet operator sequences, operatively linked to a heterologous second polypeptide which inhibits transcription in eukaryotic cells.

Claim 46
A kit comprising a carrier means having in close confinement therein at least two container means comprising:

a) a first container means containing a first nucleic acid encoding a fusion protein which inhibits transcription in eukaryotic cells, the fusion protein comprising
a first polypeptide which binds to tet operator sequences either

i. in the presence but not the absence of tetracycline or a tetracycline analogue or

ii. in the absence but not the presence of tetracycline or a tetracycline analogue,
operatively linked to a heterologous second polypeptide which inhibits transcription in eukaryotic cells, or
a eukaryotic cell line into which said first nucleic acid has been stably introduced; and

b) a second container means containing a second nucleic acid comprising a cloning site for introduction of a nucleotide sequence to be transcribed operatively linked to at least one tet operator sequence.

Claim 1
A method for regulating expression of a tet operator-linked gene in a cell of a subject, comprising:

a) introducing into the cell a nucleic acid molecule encoding a fusion protein which inhibits transcription in eukaryotic cells, the fusion protein comprising
a first polypeptide which binds to a tet operator sequence, operatively linked to a heterologous second polypeptide which inhibits transcription in eukaryotic cells; and
b) modulating the concentration of a tetracycline, or analogue thereof, in the subject.

Claim 17
A method for regulating expression of a gene in a cell of a subject, comprising:

a) obtaining the cell from the subject;
b) introducing into the cell a first nucleic acid molecule which operatively links a gene to at least one tet operator sequence;
c) introducing into the cell a second nucleic acid molecule encoding a fusion protein which inhibits transcription, the fusion protein comprising
a first polypeptide which binds to a tet operator sequence, operatively linked to a second polypeptide which inhibits transcription in eukaryotic cells, to form a modified cell;
d) administering the modified cell to the subject; and
e) modulating the concentration of a tetracycline, or analogue thereof, in the subject.

Claim 1
A method for regulating expression of a tet operator-linked gene in a cell of a subject, comprising:

a) introducing into the cell a nucleic acid molecule encoding a tetracycline-controllable transactivator (tTA), the tTA comprising
a Tet repressor operably linked to a polypeptide which directly or indirectly activates transcription in eucaryotic cells; and
b) modulating the concentration of a tetracycline, or analogue thereof, in the subject.

Claim 10
A method for regulating expression of a gene in a cell of a subject, comprising:

a) obtaining the cell from the subject;
b) introducing into the cell a first nucleic acid molecule which operatively links a gene to at least one tet operator sequence;
c) introducing into the cell a second nucleic acid molecule encoding a te tracycline-controllable transactivator (tTA), the tTA comprising
a Tet repressor operably linked to a polypeptide which directly or indirectly activates transcription in eucaryotic cells, to form a modified cell;
d) administering the modified cell to the subject; and
e) modulating the concentration of a tetracycline, or analogue thereof, in the subject.

Claim 1
A method for regulating expression of a tet operator-linked gene in a cell of a subject, comprising:

a) introducing into the cell a nucleic acid molecule encoding a fusion protein which activates transcription, the fusion protein comprising
a first polypeptide which binds to a tet operator sequence in the presence of tetracycline or a tetracycline analogue operatively linked to
a second polypeptide which activates transcription in eukaryotic cells; and
b) modulating the concentration of a tetracycline, or analogue thereof, in the subject, such that expression of a tet operator-linked gene in a cell of the subject is regulated.

Claim 16
A method for regulating expression of a gene in a cell of a subject, comprising:

a) obtaining the cell from the subject;
b) introducing into the cell a first nucleic acid molecule which operatively links a gene to at least one tet operator sequence;
c) introducing into the cell a second nucleic acid molecule encoding a fusion protein which activates transcription, the fusion protein comprising
a first polypeptide which binds to a tet operator sequence in the presence of tetracycline or a tetracycline analogue operatively linked to
a second polypeptide which activates transcription in eukaryotic cells, to form a modified cell;
d) administering the modified cell to the subject; and
e) modulating the concentration of a tetracycline, or analogue thereof, in the subject such that expression of the gene which is operatively linked to at least one tet operator sequence is regulated in a cell of the subject.

Claim 28
An isolated recombinant vector for coordinate, bidirectional transcription of a first and a second nucleotide sequence,
the vector comprising a nucleotide sequence comprising in a 5' to 3' direction:

a) a first cloning site for introduction of a first nucleotide sequence to be transcribed, operatively linked to
b) at least one tet operator sequence, operatively linked to
c) a second cloning site for introduction of a second nucleotide sequence transcribed,
the vector further comprising additional regulatory sequences such that the vector is sufficient for use in eukaryotic cells,
wherein transcription of a first and second nucleotide sequence introduced into the vector proceeds in opposite directions relative to the at least one tet operator sequence.

Claim 35
An isolated nucleic acid composition comprising at least one recombinant vector for independent regulation of transcription of a first and a second nucleotide sequence, the nucleic acid composition comprising nucleotide sequences comprising:

a) a first cloning site for introduction of a first nucleotide sequence to be transcribed, operatively linked to at least one tet operator sequence of a first class type; and
b) a second cloning site for introduction of a second nucleotide sequence to be transcribed, operatively linked to at least one tet operator sequence of a second class type.

Claim 1
A transgenic mouse having a transgene integrated into the genome of the mouse and also having a tet operator-linked gene in the genome of the mouse, wherein:

a)the transgene comprises a transcriptional regulatory element functional in cells of the mouse operatively linked to a polynucleotide sequence encoding a tetracycline-controllable transactivator fusion protein (tTA),
b) said fusion protein comprises a Tet repressor operably linked to a polypeptide which directly or indirectly activates transcription of said tet operator-linked gene in eucaryotic cells,
c) said tet operator-linked gene confers a detectable and functional phenotype on the mouse when expressed in cells of the mouse,
d) said transgene is expressed in cells of the mouse at a level sufficient to produce amounts of said fusion protein that are sufficient to activate transcription of the tet operator-linked gene; and
e) in the absence of tetracycline or a tetracycline analogue in the mouse, said fusion protein binds to the tet operator-linked gene and activates transcription of the tet operator linked gene such that the tet operator-linked gene is expressed at a level sufficient to confer the detectable and functional phenotype on the mouse,

wherein the level of expression of tet operator-linked gene can be down modulated by administering tetracycline or a tetracycline analogue to the mouse.

Claim 13
A transgenic mouse having a transgene integrated into the genome of the mouse, wherein:

a) the transgene comprises a polynucleotide sequence encoding a tet racycline-controllable transactivator fusion protein (tTA) and a tTA-responsive promoter,
b) said fusion protein comprises a Tet repressor operably linked to a polypeptide which directly or indirectly activates transcription of a gene of interest in eucaryotic cells,
c) the transgene is integrated by homologous recombination at a predetermined location within a said gene of interest within cells of the mouse such that expression of the fusion protein is controlled by 5' regulatory elements of the gene of interest and expression of the gene of interest is controlled by the tTA-responsive promoter, expression of the gene of interest confers a detectable and functional phenotype on the mouse,
d) said transgene is expressed in cells of the mouse at a level sufficient to produce amounts of said fusion protein that are sufficient to activate transcription of the gene of interest linked to the tTA-responsive promoter, and
e) in the absence of tetracycline or a tetracycline analogue in the mouse, said fusion protein binds to the tTA-responsive promoter and activates transcription of the gene of interest such that the gene of interest is expressed at a level sufficient to confer the detectable and functional phenotype on the mouse,
f) wherein the level of expression of the gene of interest can be down modulated by administering tetracycline or a tetracycline analogue to the mouse.

Claim 20
A transgenic mouse having a transgene integrated into the genome of the mouse, wherein:

a) the transgene comprises a transcriptional regulatory element functional in cells of the mouse operatively linked to a polynucleotide sequence encoding a tetracycline-controllable transactivator fusion protein (tTA),
b) said fusion protein comprises a Tet repressor operably linked to a polypeptide which directly or indirectly activates transcription of a tet operator-linked gene in eucaryotic cells, and
c) said fusion protein is expressed in cells of the mouse.

Claim 1
A transgenic mouse having a transgene integrated into the genome of the mouse and also having a tet operator-linked gene in the genome of the mouse, wherein:

a) the transgene comprises a transcriptional regulatory element functional in cells of the mouse operatively linked to a polynucleotide sequence encoding a fusion protein which inhibits transcription of said tet operator linked gene,
b) said fusion protein comprises a first polypeptide that is a Tet repressor operably linked to a heterologous second polypeptide which inhibits transcription of said tet operator-linked gene in eucaryotic cells,
c) said tet operator-linked gene confers a detectable and functional phenotype on the mouse when expressed in cells of the mouse,
d) said transgene is expressed in cells of the mouse at a level sufficient to produce amounts of said fusion protein that are sufficient to inhibit transcription of the tet operator-linked gene; and
e) in the absence of tetracycline or a tetracycline analogue in the mouse, said fusion protein binds to the tet operator-linked gene and inhibits transcription of the tet operator linked gene,
f) wherein the level of expression of the tet operator-linked gene can be upregulated by administering tetracycline or a tetracycline analogue to the mouse.

Claim 2
A transgenic mouse having a transgene integrated into the genome of the mouse and also having a tet operator-linked gene in the genome of the mouse, wherein:

a) the transgene comprises a transcriptional regulatory element functional in cells of the mouse operatively linked to a polynucleotide sequence encoding a fusion protein which inhibits transcription of said tet operator linked gene,
b) said fusion protein comprises a first polypeptide that is a mutated Tet repressor that binds to tet operator sequences in the presence, but not the absence, of tetracycline or a tetracycline analogue, operably linked to a heterologous second polypeptide which inhibits transcription of
c) said tet operator-linked gene in eucaryotic cells,
d) said tet operator-linked gene confers a detectable and functional phenotype on the mouse when expressed in cells of the mouse,
e) said transgene is expressed in cells of the mouse at a level sufficient to produce amounts of said fusion protein that are sufficient to inhibit transcription of the tet operator-linked gene; and
f) in the presence of tetracycline or a tetracycline analogue in the mouse, said fusion protein binds to the tet operator-linked gene and inhibits transcription of the tet operator linked gene,
g) wherein the level of expression of the tet operator-linked gene can be upregulated by depleting tetracycline or a tetracycline analogue from the mouse.

Claim 3
A transgenic mouse having a transgene integrated into the genome of the mouse, wherein:

a) the transgene comprises a transcriptional regulatory element functional in cells of the mouse operatively linked to a polynucleotide sequence encoding a fusion protein which inhibits transcription of a tet operator-linked gene,
b) the fusion protein comprising a first polypeptide that is a Tet repressor or, a mutated Tet repressor that binds to a tet operator sequence, operatively linked to a second polypeptide which inhibits transcription in eukaryotic cells, and
c) said fusion protein is expressed in cells of the mouse.

Claim 1
A transgenic mouse having a transgene integrated into the genome of the mouse and also having a tet operator-linked gene in the genome of the mouse, wherein:

a) the transgene comprises a transcriptional regulatory element functional in cells of the mouse operatively linked to a polynucleotide sequence encoding a fusion protein which activates transcription of said tet operator-linked gene,
b) the fusion protein comprises a first polypeptide which is a mutated Tet repressor that binds to a tet operator sequence in the presence of tetracycline or a tetracycline analogue operatively linked to a second polypeptide which activates transcription in eukaryotic cells,
c) said tet operator-linked gene confers a detectable and functional phenotype on the mouse when expressed in cells of the mouse,
d) said transgene is expressed in cells of the mouse at a level sufficient to produce amounts of said fusion protein that are sufficient to activate transcription of the tet operator-linked gene; and
e) in the presence of tetracycline or a tetracycline analogue in the mouse, said fusion protein binds to the tet operator-linked gene and activ ates transcription of the tet operator-linked gene such that the tet operator-linked gene is expressed at a level sufficient to confer the detectable and functional phenotype on the mouse,
f) wherein the level of expression of the tet operator-linked gene cap be down modulated by depleting tetracycline or a tetracycline analogue from the mouse.

Claim 35
A transgenic mouse having a transgene integrated into the genome of the mouse, wherein:

a) the transgene comprises a transcriptional regulatory element functional in cells of the mouse operatively linked to a polynucleotide sequence encoding a fusion protein which activates transcription of a tet operator-linked gene,
b) the fusion protein comprising a first polypeptide which is a mutated Tet repressor that binds to a tet operator sequence in the presence of tetracycline or a tetracycline analogue operatively linked to a second po lypeptide which activates transcription in eukaryotic cells, and
c) said fusion protein is expressed in cells of the mouse.

Claim 1
A method for producing a transgenic mouse, comprising:

a) introducing into a fertilized oocyte of a mouse a DNA molecule encoding a tetracycline-controllable transactivator (tTA),
the tTA comprising a prokaryotic Tet repressor operably linked to a polypeptide which directly or indirectly activates transcription in eucaryotic cells;
b) implanting the fertilized oocyte in a pseudopregnant foster mother; and
c) allowing the fertilized oocyte to develop into a transgenic mouse to thereby produce the transgenic mouse, wherein said tTA is expressed in cells of the mouse at a level sufficient to transactivate a tet operator-linked gene.

Claim 2
A method for producing a transgenic mouse having a transgene encoding a tetracycline-controllable transactivator (tTA) integrated at a predetermined location within chromosomal DNA of cells of the mouse, comprising:

a) introducing into a population of embryonic stem cells of a mouse a DNA molecule encoding a tTA,
the tTA comprising a prokaryotic Tet repressor operably linked to a polypeptide which directly or indirectly activates transcription in eucaryotic cells,
the DNA molecule comprising a polynucleotide sequence encoding the tTA flanked at 5' and 3' ends by additional polynucleotide sequences of sufficient length for homologous recombination between the DNA molecule and a second target DNA molecule at a predetermined location within chromosomal DNA of cells of the mouse, under conditions suitable for homologous recombination between the DNA molecule encoding the tTA and chromosomal DNA within the cell;
b) selecting an embryonic stem cell in which DNA encoding the tTA has integrated at a predetermined location within the chromosomal DNA of the cell;
c) implanting the embryonic stem cell into a blastocyst;
d) implanting the blastocyst into a pseudopregnant foster mother; and
e) allowing the blastocyst to develop into a transgenic mouse to thereby produce the transgenic mouse,
wherein said tTA is expressed in cells of the mouse at a level sufficient to transactivate a tet operator-linked gene.

Claim 3
A method for producing a transgenic mouse having a transgene encoding a tetrac ycline-controllable transactivator (tTA) and a tTA-responsive promoter integrated at a predetermined location within a gene of interest in cells of the mouse, comprising:

a) introducing into a population of embryonic stem cells of a mouse a DNA molecule encoding a tTA, the DNA molecule comprising:

1. a first polynucleotide sequence comprising a 5' flanking regulatory region of the gene of interest, operably linked to;
2. a second polynucleotide sequence encoding a tTA, the tTA comprising a prokaryotic Tet repressor operably linked to a polypeptide which directly or indirectly activates transcription in eucaryotic cells; and
3. a third polynucleotide sequence comprising a tTA-responsive promoter, operably linked to;
4. a fourth polynucleotide sequence comprising at least a portion of a coding region of the gene of interest;
5. wherein the first and fourth polynucleotide sequences are of sufficient length for homologous recombination between the DNA molecule and the gene of interest such that expression of the tTA is controlled by 5' regulatory elements of the gene of interest and expression of the gene of interest is controlled by the tTA-responsive promoter, under conditions suitable for homologous recombination between the DNA molecule encoding the tTA and the gene of interest within the cell;

b) selecting an embryonic stem cell in which DNA encoding the tTA has integrated at a predetermined location within the gene of interest in the cell;
c) implanting the embryonic stem cell into a blastocyst;
d) implanting the blastocyst into a pseudopregnant foster mother; and
e) allowing the blastocyst to develop into a transgenic mouse to thereby produc e the transgenic mouse,
wherein said gene of interest confers a detectable and functional phenotype on the mouse when expressed in cells of the transgenic mouse, said tTA is expressed in cells of the transgenic mouse at a level sufficient to activate transcription of the gene of interest; and
in the absence of tetracycline or a tetracycline analogue in the mouse, said tTA binds to the tTA responsive promoter operably linked to the gene of interest and activates transcription of the gene of interest such that the gene of interest is expressed at a level sufficient to confer the detectable and functional phenotype on the mouse,
wherein the level of expression of the gene of interest can be down modulated by administering tetracycline or a tetracycline analogue to the mouse.

Claim 1
A fusion protein which activates transcription comprising:
a first polypeptide which binds to a tet operator sequence in the presence of tetracycline or a tetracycline analogue operatively linked
to
a second polypeptide which activates transcription in eukaryotic cells.

Claim 1
A fusion protein which inhibits transcription in eukaryotic cells, the fusion protein comprising:
a first polypeptide which binds to tet operator sequences, operatively linked to
a heterologous second polypeptide which inhibits transcription in eukaryotic cells.

Claim 1
A transgenic plant having a transgene integrated into the genome of the plant and also having a tet operator-linked gene in the genome of the plant, wherein:

a) the transgene comprises:
a transcriptional regulatory element functional in cells of the plant operatively linked to a polynucleotide sequence encoding a fusion protein which activates transcription of said tet operator-linked gene,

the fusion protein comprises
a) a first polypeptide which is a mutated Tet repressor that binds to a te t operator sequence in the presence of tetracycline or a tetracycline analogue operatively linked to
b) a second polypeptide which activates transcription in eukaryotic cells,

b) said tet operator-linked gene confers a detectable and functional phenotype on the plant when expressed in cells of the plant,
c) said transgene is expressed in cells of the plant at a level sufficient to produce amounts of said fusion protein that are sufficient to activate transcription of the tet operator-linked gene; and
d) in the presence of tetracycline or a tetracycline analogue in the plant, said fusion protein binds to the tet operator-linked gene and activates transcription of the tet operator-linked gene such that the tet operator-linked gene is expressed at a level sufficient to confer the detectable and functional phenotype on the plant,
wherein the level of expression of the tet operator-linked gene can be down modulated by depleting tetracycline or a tetracycline analogue from the plant.

Claim 2
A transgenic plant having a transgene integrated into the genome of the plant, wherein:

a) the transgene comprises
a transcriptional regulatory element functional in cells of the plant operatively linked to a polynucleotide sequence encoding a fusion protein which activates transcription of a tet operator linked gene,

the fusion protein comprising
a) a first polypeptide which is a mutated Tet repressor that binds to a tet operator sequence in the presence of tetracycline or a tetracycline analogue operatively linked to
b) a second polypeptide which activates transcription in eukaryotic cells, and
said fusion protein is expressed in cells of the plant.

Claim 3
A transgenic plant having a transgene integrated into the genome of the plant and also having a tet operator-linked gene in the genome of the plant, wherein:

the transgene comprises
a transcriptional regulatory element functional in cells of the plant operatively linked to a polynucleotide sequence encoding a fusion protein which inhibits transcription of said tet operator linked gene,
the fusion protein comprises
a first polypeptide which is a mutated Tet repressor that binds to a tet operator sequence in the presence of tetracycline or a tetracycline analogue operatively linked to
a second polypeptide which inhibits transcription in eukaryotic cells,

said tet operator-linked gene confers a detectable and functional phenotype on the plant when expressed in cells of the plant,
said transgene is expressed in cells of the plant at a level sufficient to produce amounts of said fusion protein that are sufficient to inhibit transcription of the tet operator-linked gene; and
in the presence of tetracycline or a tetracycline analogue in the plant, said fusion protein binds to the tet operator-linked gene and inhibits transcription of the tet operator-linked gene, wherein the level of expression of the tet operator-linked gene can be upregulated by depleting tetracycline or a tetracycline analogue from the plant.

Claim 4
A transgenic plant having a transgene integrated into the genome of the plant, wherein: the transgene comprises a transcriptional regulatory element functional in cells of the plant operatively linked to a polynucleotide sequence encoding a fusion protein which inhibits transcription of a tet operator linked gene, the fusion protein comprising a first polypeptide which is a mutated Tet repressor that binds to a tet operator sequence in the presence of tetracycline or a tetracycline analogue operatively linked to a second polypeptide which inhibits transcription in eukaryotic cells, and said fusion protein is expressed in cells of the plant.

Claim 5
A transgenic plant having a transgene integrated into the genome of the plant and also having a tet operator-linked gene in the genome of the plant, wherein: the transgene comprises a transcriptional regulatory element functional in cel ls of the plant operatively linked to a polynucleotide sequence encoding a fusion protein which inhibits transcription of said tet operator linked gene, said fusion protein comprises a first polypeptide that is a Tet repressor, operably linked to a heterologous second polypeptide which inhibits transcription of said tet operator-linked gene in eukaryotic cells, said tet operator-linked gene confers a detectable and functional phenotype on the plant when expressed in cells of the plant, said transgene is expressed in cells of the plant at a level sufficient to produce amounts of said fusion protein that are sufficient to inhibit transcription of the tet operator-linked gene; and in the absence of tetracycline or a tetracycline analogue in the plant, said fusion protein binds to the tet operator-linked gene and inhibits transcription of the tet operator linked gene, wherein the level of expression of the tet operator-linked gene can be upregulated by administering tetracycline or a tetracycline analogue to the plant.

Claim 6
A transgenic plant having a transgene integrated into the genome of the plant, wherein: the transgene comprises a transcriptional regulatory element functional in cells of the plant operatively linked to a polynucleotide sequence encoding a fusion protein which inhibits transcription of a tet operator linked gene, the fusion protein comprising a first polypeptide which is a Tet repressor, operatively linked to a second polypeptide which inhibits transcription in eukaryotic cells, and said fusion protein is expressed in cells of the plant.

Claim 1
A transgenic plant having a transgene integrated into the genome of the plant and also having a tet operator-linked gene in the genome of the plant, wherein: the transgene comprises a transcriptional regulatory element functional in c ells of the plant operatively linked to a polynucleotide sequence encoding a fusion protein which activates transcription of said tet operator linked gene, the fusion protein comprises a first polypeptide which is a Tet repressor operatively linked to a second polypeptide which directly or indirectly activates transcription in plant cells, said tet operator-linked gene confers a detectable and functional phenotype on the plant when expressed in cells of the plant, said transgene is expressed in cells of the plant at a level sufficient to produce amounts of said fusion protein that are sufficient to activate transcription of the tet operator-linked gene; and in the absence of tetracycline or a tetracycline analogue in the plant, said fusion protein binds to the tet operator-linked gene and activates transcription of the tet operator linked gene such that the tet operator-linked gene is expressed at a level sufficient to confer the detectable and functional phenotype on the plant, wherein the level of expression of the tet operator-linked gene can be down modulated by administering tetracycline or a tetracycline analogue to the pl ant.

Claim 7
A transgenic plant having a transgene integrated into the genome of he plant, wherein: the transgene comprises a transcriptional regulatory element fun ctional in cells of the plant operatively linked to a polynucleotide sequence encoding a fusion protein which activates transcription of a tet operator linked gene, the fusion protein comprising a first polypeptide which is a Tet repressor, operatively linked to a second polypeptide which directly or indirectly activates transcription in plant cells, and said fusion protein is expressed in cells of the plant.

Claim 1
An isolated nucleic acid encoding a fusion protein which activates transcription,
the fusion protein comprising a first polypeptide which binds to a tet operator sequence in the presence, but not the absence, of tetracycline or a tetracycline analogue operatively linked to a second polypeptide which activates transcription in eukaryotic cells.

Claim 12
A fusion protein which activates transcription comprising a first polypeptide which binds to a tet operator sequence in the presence, but not the absence, of tetracycline or a tetracycline analogue operatively linked to a second polypeptide which activates transcription in eukaryotic cells .

Claim 35
An isolated nucleic acid encoding a fusion protein which inhibits transcription in eukaryotic cells,
the fusion protein comprising a first polypeptide which binds to a tet operator sequence operatively linked to a heterologous second polypeptide which inhibits transcription in eukaryotic cells.

Claim 51
A fusion protein which inhibits transcription in eukaryotic cells, comprising a first polypeptide which binds to a tet operator sequence operatively linked to a heterologous second polypeptide which inhibits transcription in eukaryotic cells.

Claim 73
A host cell comprising:

a) a first nucleic acid encoding a first fusion protein which activates transcription,
the first fusion protein comprising a first polypeptide which binds to a tet operator sequence operatively linked to a second polypeptide which activates transcription in eukaryotic cells;
b) a second nucleic acid encoding a second fusion protein which inhibits transcription,
the second fusion protein comprising a third polypeptide which binds to a tet operator sequence operatively linked to a fourth polypeptide which inhibits transcription in eukaryotic cells; and
c) a third nucleic acid molecule comprising a nucleotide sequence to be transcribed operatively linked to at least one tet operator sequence.

Claim 76
A non-human transgenic organism comprising:

a) a first transgene encoding a first fusion protein which activates transcription,
the first fusion protein comprising a first polypeptide which binds to a tet operator sequence operatively linked to a second polypeptide which activates transcription in eukaryotic cells;
b) a second transgene encoding a second fusion protein which inhibits transcription,
the second fusion protein comprising a third polypeptide which binds to a tet operator sequence operatively linked to a fourth polypeptide which inhibits transcription in eukaryotic cells; and
c) a third transgene comprising a nucleotide sequence to be transcribed operatively linked to at least one tet operator sequence.

Claim 79
A recombinant vector for coordinate, bidirectional transcription of a first and a second nucleotide sequence to be transcribed,
the vector comprising a nucleotide sequence comprising in a 5' to 3' direction:
a first cloning site for introduction of a first nucleotide sequence to be transcribed,
which is operatively linked to at least one tet operator sequence,
which is operatively linked to a second cloning site for introduction of a second nucleotide sequence to be transcribed,
wherein transcription of the first and second nucleotide sequence introduced into the vector proceeds in opposite directions relative to the at least one tet operator sequence.

Claim 83
A composition of matter comprising at least one recombinant vector for independent regulation of transcription of a first and a second nucleotide sequence to be transcribed, the at least one vector comprising a nucleotide sequence comprising:

a) a first cloning site for introduction of a first nucleotide sequence to be transcribed, operatively linked to at least one tet operator sequence of a first class type; and
b) a second cloning site for introduction of a second nucleotide sequence to be transcribed, operatively linked to at least one tet operator sequence of a second class type.

Claim 87
A kit comprising a carrier means having in close confinement therein at least two container means comprising:

a) a first container means containing a first nucleic acid encoding a fusion protein which activates transcription,
the fusion protein comprising a first polypeptide which binds to a tet operator sequence in the presence of tetracycline or a tetracycline analogue operatively linked to a second polypeptide which activates transcription in eukaryotic cells; and
b) a second container means containing a second nucleic acid comprising a cloning site for introduction of a nucleotide sequence to be transcribed operatively linked to at least one tet operator sequence.

Claim 1
An isolated nucleic acid encoding a fusion protein which inhibits transcription in eukaryotic cells,
the fusion protein comprising:

a) a first polypeptide which is a Tet repressor that binds to a tet operator sequence in the absence but not the presence of tetracycline or a tetracycline analogue operatively linked to
b) a heterologous second polypeptide which inhibits transcription in eukaryotic cells.

Claim 1
An expression vector adapted for replication in an animal cell comprising a glucocorticoid responsive promoter,
said promoter comprising

a) a plurality of at least 5 glucocorticoid response elements (GREs),
b) a viral or mammalian TATA box, and
c) a viral or mammalian initiator element with a transcriptional initiator site located from 20 to 50 bases from said TATA box,

said promoter lacking upstream elements which bind nuclear factor I, and
said vector further comprising a restriction endonuclease site downstream from said promoter for insertion of DNA to be expressed from said promoter; wherein said DNA is expressed from said vector in an animal cell.

Claim 10
A promoter consisting of:

a) a plurality of at least five glucocorticoid response elements (GREs),
b) a TATA box, and
c) an initiator site containing a transcriptional initiator site located from 20 to 50 bases from said TATA box,

said promoter lacking upstream elements which bind nuclear factor I,
wherein said promoter is responsive to ligand-bound glucocorticoid, progesterone, androgen or mineralocorticoid receptor when transiently transfected into cells, when stably integrated within a genome, or when stably propagated in an episomal vector.

Claim 1
A method for selecting transgenic plants comprising a silent selectable marker wherein said method comprises the steps of:

a) transforming a plant cell with a vector wherein said vector comprises DNA encoding a regulatory region of an estrogen receptor and further wherein said vector comprises a gene which promotes shoot formation, wherein said gene is under the control of an inducible promoter;
b) growing said plant cells in the absence of a plant hormone but in the presence of an inducer of said inducible promoter; and
c) excising shoots which develop, wherein said shoots can grow into transgenic plants when grown in the absence of said inducer.

Claim 14
A method for inducing plant somatic embryo formation comprising the steps of:
a) transforming a plant cell with a vector encoding a gene which promotes somatic embryogenesis, wherein said gene is under the control of an inducible promoter; and
b) growing said plant cells in the absence of a plant hormone but in the presence of an inducer of said inducible promoter, wherein somatic embryos will develop.

Claim 26
A method for selecting transgenic plants wherein said method comprises growing a transgenic plant, comprising an antibiotic resistance gene under the control of a promoter comprising DNA encoding a regulatory domain of an estrogen receptor inducible by 17-estradiol or 4-hydroxyl tamoxifen, in the presence of an antibi otic, wherein said antibiotic is one to which resistance is conferred by said antibiotic resistance gene, and in the presence of 17- estradiol or 4-hydroxyl tamoxifen.

Claim 27
A method for selecting transgenic plants wherein said method comprises growing a transgenic plant, comprising a herbicide resistance gene wherein said herbicide resistance gene is under the control of a promoter comprising DNA encoding a regulatory domain of an estrogen receptor inducible by 17-estradiol or 4-hydroxyl tamoxifen, in the presence of a herbicide, wherein said herbicide is one to which resistance is conferred by said herbicide resistance gene, and in the presence of 17-estradiol or 4-hydroxyl tamoxifen.

Claim 1
A method for selecting transgenic lettuce plants comprising a silent selectable marker wherein said method comprises the steps of : a) transforming lettuce root cells with a vector wherein said vector comprises a gene selected from the group consisting of an ipt gene, a CKI1 gene, a gene from the knotted family, and a gene the expression of which is capable of promoting shoot regeneration, wherein said gene is under the control of an inducible promoter ; b) growing said lettuce root cells to allow shoot development ; and c) excising shoots which develop from plants having a shooty phenotype, wherein said shoots can grow into normal transgenic plants when grown in the absence of said inducer.

Claim 11
A vector comprising a chemically inducible promoter wherein said vector comprises DNA encoding an estrogen receptor.

Claim 25
A vector comprising:

i) a constitutive promoter,
ii) DNA encoding a DNA binding domain of bacterial repressor LexA,
iii) DNA encoding a transactivating domain of VP16,
iv) DNA encoding an estrogen receptor, and
v) one or more LexA binding sites.

Claim 26
A nucleic acid comprising a chemically inducible promoter wherein said nucleic acid further comprises DNA encoding an estrogen receptor.

Claim 41
A nucleic acid comprising i) a constitutive promoter, ii) DNA encoding a DNA binding domain of bacterial repressor LexA, iii) DNA encoding a transactivating domain of VP 16, iv) DNA encoding an estrogen receptor, and v) one or more LexA binding sites.

Claim 42
A transgenic lettuce plant or transgenic lettuce plant cell comprising a vector wherein said vector comprises a chemically inducible promoter.

Claim 43
A transgenic plant or transgenic plant cell comprising a vector wherein said vector comprises a chemically inducible promoter which can be induced by an estrogen.

Claim 50
A method for making a transgenic plant display a fluorescent design, a word or words wherein said method comprises the steps of : a) preparing a transgenic plant which comprises a luciferase gene under the control of a chemically inducible promoter which is controlled by an estrogen ; and b) placing a chemical which induces said chemically inducible promoter onto said transgenic plant in the pattern of the design, word or words which are desired ; whereby said plant will produce luciferase and will fluoresce in the pattern in which the chemically inducible promoter was placed onto said transgenic plant

Claim 51
A transgenic lettuce plant comprising an antibiotic resistance gene wherein said antibiotic resistance gene is under the control of an inducible promoter.

Claim 52
A transgenic plant comprising an antibiotic resistance gene wherein said antibiotic resistance gene is under the control of an inducible promoter, wherein said inducible promoter comprises DNA encoding a regulatory domain of an estrogen receptor.

Claim 57
A transgenic plant comprising a herbicide resistance gene wherein said herb icide resistance gene is under the control of an inducible promoter, wherein said inducible promoter comprises DNA encoding a regulatory domain of an estrogen receptor

Claim 61
An organism or a cell comprising a gene wherein a natural promoter of said gene is lacking or inoperative and said gene is under the control of a transgenic inducible promoter

Claim 74
A method to screen for mutations in a gene of an organism or cell comprising:

a) preparing an organism or a cell wherein a natural promoter of said gene is lacking or inoperative and said gene is under the control of a transgenic inducible promoter; and
b) growing said organism or cell

Claim 1
An isolated or synthetic DNA sequence encoding a polypeptide selected from the group consisting of:

a) the Heliothis virescens ecdysone steroid receptor shown in SEQ ID NO: 5;
b) the transactivation domain of the Heliothis virescens ecdysone steroid receptor shown in amino acids 1-162 of SEQ ID NO: 5;
c) the DNA binding domain of the Heliothis virescens ecdysone steroid receptor shown in amino acids 163-228 of SEQ ID NO: 5;
d) the hinge domain of the Heliothis virescens ecdysone steroid receptor shown in amino acids 229-326 of SEQ ID NO: 5;
e) the ligand binding domain of the Heliothis virescens ecdysone steroid receptor shown in amino acids 327-545 of SEQ ID NO: 5;
f) the carboxy terminus of the Heliothis virescens ecdysone steroid receptor shown in amino acids 546-577 of SEQ ID NO: 5; and
g) the hinge and ligand binding domains of the Spodoptera exigua ecdysone steroid receptor shown in SEQ ID NO: 7.

Claim 1
DNA comprising the sequence shown in Seq. ID No. 2, or a DNA sequence which hybridizes to said DNA sequence under high stringency conditions.

Claim 2
DNA comprising the sequence shown in Seq. ID No.3, or a DNA sequence which hybridizes to said DNA sequence under high stringency conditions.

Claim 3
DNA comprising the sequence shown in Seq. ID No. 4, or a DNA sequence which hybridizes to said DNA sequence under high stringency conditions

Claim 11
A polypeptide comprising the Heliothis ecdysone receptor or a func tionally active fragment thereof, wherein the polypeptide is substantially free from other proteins with which is ordinarily associated, and which is coded for by the DNA of any one of the preceding claims

Claim 12
A polypeptide comprising the amino acid sequence shown in Seq. ID. No. 4 or any allelic variant or derivative thereof.

Claim 13
A polypeptide comprising part of the amino acid sequence shown in Seq. ID. No. 4 or any allelic variant or derivative thereof, which sequence provides the Heliothis ecdysone receptor ligand binding domain.

Claim 14
A polypeptide comprising part of the amino acid sequence shown in Seq. ID. No. 4 or any allelic variant or derivative thereof, which sequence provides the Heliothis ecdysone receptor DNA binding domain.

Claim 15
A polypeptide comprising part of the amino acid sequence shown in Seq. ID. No. 4 or any allelic variant or derivative thereof, which sequence provides the Heliothis ecdysone receptor transactivation domain.

Claim 16
A polypeptide comprising part of the amino acid sequence shown in Seq. ID. No. 4 or any allelic variant or derivative thereof, which sequence provides the Heliothis ecdysone receptor hinge domain.

Claim 17
A polypeptide comprising part of the amino acid sequence shown in Seq. ID. No. 4 or any allelic variant or derivative thereof, which sequence provides the Heliothis ecdysone receptor carboxy terminal domain.

Claim 19
DNA comprising the sequence shown in Seq. ID. No. 6, or a DNA sequence which hybridizes to said DNA sequence under high stringency conditions.

Claim 1
DNA comprising the sequence shown in Seq ID No. 2.

Claim 2
DNA comprising the sequence shown in Seq ID No. 3.

Claim 3
DNA comprising the sequence shown in Seq ID No. 4.

Claim 4
DNA comprising a sequence which shows 60% or more homology with the sequence shown in Seq ID No 1, 2 or 3.

Claim 6
DNA which hybridizes to the sequence shown in Seq. ID No. 2, 3 or 4, and which codes for at least part of the Heliothis ecdysone receptor.

Claim 8
DNA comprising part of the sequence shown in Seq ID No. 2, and which codes for at least part of the Heliothis ecdysone receptor ligand binding domain.

Claim 9
DNA comprising part of the sequence shown in Seq ID No. 3, and which codes for at least part of the Heliothis ecdysone receptor ligand binding domain.

Claim 10
DNA comprising part of the sequence shown in Seq ID No. 4, and which codes for at least part of the Heliothis ecdysone receptor ligand binding domain.

Claim 15
DNA comprising part of the sequence shown in Seq ID No. 2, and which codes for at least part of the Heliothis ecdysone receptor DNA binding domain.

Claim 16
DNA comprising part of the sequence shown in Seq ID No. 3, and which codes for at least part of the Heliothis ecdysone receptor DNA binding domain.

Claim 17
DNA comprising part of the sequence shown in Seq ID No. 4, and which codes for at least part of the Heliothis ecdysone receptor DNA binding domain.

Claim 22
DNA comprising part of the sequence shown in Seq ID No. 2, and which codes for at least part of the Heliothis ecdysone receptor transactivation domain.

Claim 23
DNA comprising part of the sequence shown in Seq ID No. 3, and which codes for at least part of the Heliothis ecdysone receptor transactivation domain.

Claim 24
DNA comprising part of the sequence shown in Seq ID No. 4, and which codes for at least part of the Heliothis ecdysone receptor transactivation domain.

Claim 29
DNA comprising part of the sequence shown in Seq ID No. 2, and which codes for at least part of the Heliothis ecdysone receptor hinge domain.

Claim 30
DNA comprising part of the sequence shown in Seq ID No. 3, and which codes for at least part of the Heliothis ecdysone receptor hinge domain.

Claim 31
DNA comprising part of the sequence shown in Seq ID No. 4, and which codes for at least part of the Heliothis ecdysone receptor hinge domain.

Claim 36
DNA having part of the sequence shown in Seq ID No. 2, and which codes for at least part of the Heliothis ecdysone receptor carboxy terminal region.

Claim 37
DNA having part of the sequence shown in Seq ID No. 3, and. which codes for at least part of the Heliothis ecdysone receptor carboxy terminal region.

Claim 38
DNA having part of the sequence shown in Seq ID No. 4, and which codes for at least part of the Heliothis ecdysone receptor carboxy terminal region.

Claim 44
A polypeptide comprising the amino acid sequence shown in Seq ID No. 4 or any allelic variant or derivative thereof.

Claim 45
A polypeptide comprising part of the amino acid sequence shown in Seq ID No. 4 or any allelic variant or derivative thereof, which sequence provides the Heliothis ecdysone receptor ligand binding domain.

Claim 46
A polypeptide comprising part of the amino acid sequence shown in Seq ID No. 4 or any allelic variant or derivative thereof, which sequence provides the Heliothis ecdysone receptor DNA binding domain.

Claim 47
A polypeptide comprising part of the amino acid sequence shown in Seq ID No. 4 or any allelic variant or derivative thereof, which sequence provides the Heliothis ecdysone receptor transactivation domain.

Claim 48
A polypeptide comprising part of the amino acid sequence shown in Seq ID No. 4 or any allelic variant or derivative thereof, which sequence provides the Heliothis ecdysone receptor hinge domain.

Claim 49
A polypeptide comprising part of the amino acid sequence shown in Seq ID No. 4 or any allelic variant or derivative thereof, which sequence provides the Heliothis ecdysone receptor carboxy terminal region.

Claim 51
DNA comprising the sequence shown in Seq ID No. 6.

Claim 52
DNA comprising a sequence which shows 60% or more homology with the sequence shown in Seq ID No. 6.

Claim 54
DNA which hybridizes to the DNA sequence shown in Seq ID No. 6 and which codes for at least part of Spodoptera ecdysone receptor.

Claim 56
DNA comprising part of the sequence shown in Seq ID No. 6, and which codes for at least part of the Spodoptera ecdysone receptor ligand binding domain.

Claim 61
DNA comprising part of the sequence shown in Seq ID No. 6, and which codes for at least part of the Spodoptera ecdysone receptor hinge domain.

Claim 1
DNA comprising the sequence shown in Seq. ID No. 2, or a sequence which shows homology thereto, or a sequence which hybridizes to the sequence shown in Seq. ID No. 2.

Claim 2
DNA comprising the sequence shown in Seq. ID No.3, or a sequence which shows homology thereto, or a sequence which hybridizes to the sequence shown in Seq. ID No. 3.

Claim 3
DNA comprising the sequence shown in Seq. ID No. 4, or a sequence which shows homology thereto, or a sequence which hybridizes to the sequence shown in Seq. ID No. 4.

Claim 14
A polypeptide comprising the amino acid sequence shown in Seq. ID. No. 4 or any allelic variant or derivative thereof.

Claim 15
A polypeptide comprising part of the amino acid sequence shown in Seq. ID. No. 4 or any allelic variant or derivative thereof, which sequence provides the Heliothis ecdysone receptor ligand binding domain.

Claim16
A polypeptide comprising part of the amino acid sequence shown in Seq. ID. No. 4 or any allelic variant or derivative thereof, which sequence provides the Heliothis ecdysone receptor DNA binding domain.

Claim 17
A polypeptide comprising part of the amino acid sequence shown in Seq. ID. No. 4 or any allelic variant or derivative thereof, which sequence provides the Heliothis ecdysone receptor transactivation domain.

Claim 18
A polypeptide comprising part of the amino acid sequence shown in Seq. ID. No. 4 or any allelic variant or derivative thereof, which sequence provides the Heliothis ecdysone receptor hinge domain.

Claim 19
A polypeptide comprising part of the amino acid sequence shown in Seq. ID. No. 4 or any allelic variant or derivative thereof, which sequence provides the Heliothis ecdysone receptor carboxy terminal domain.

Claim 21
DNA comprising the sequence shown in Seq. ID. No. 6, or a sequence which shows homology thereto, or a sequence which hybridizes to the sequence shown in Seq. ID. No. 6.

Claim 23
DNA comprising a sequence which shows homology in the range of 65% to 99% to the sequence shown in Seq. ID. No. 6.

Claim 1
An isolated nucleic acid sequence that encodes an insect protein from a Pyralidae species, wherein said nucleotide sequence is selected from the group consisting of:

a) a nucleotide sequence comprising a sequence encoding an ecdysone receptor or Ultraspiracle;
b) a nucleotide sequence comprising the sequence set forth in SEQ ID NO: 1 or 3;
c) a nucleotide sequence encoding a polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 2 or 4;
d) a nucleotide sequence that hybridizes under stringent conditions to a sequence of a), b), or c).

Claim 2
An isolated polypeptide from a Pyrilidae species, wherein said polypeptide is selected from the group consisting of.

a) a polypeptide sequence comprising an Ecdysone receptor or Ultraspiracle;
b) a polypeptide comprising an amino acid sequence set forth in 20 SEQ ID NO: 2 or 4;
c) a polypeptide encoded by a nucleotide sequence comprising the sequence set forth in SEQ ID NO: 1 or 3;
d) a polypeptide encoded by a nucleotide sequence that hybridizes under stringent conditions to a nucleotide sequence comprising the sequence set forth in SEQ ID NO: 1 or 3.

Claim 3
A method of selectively inducing gene expression of a protein of interest in a plant, said method comprising:

a) stably incorporating into the genome of said plant an expression cassette, said expression cassette comprising a promoter operably linked to a nucleotide sequence encoding an Ecdysone receptor, wherein said nucleotide sequence encoding the Ecdysone receptor is selected from the group consisting of.:

i) a nucleotide sequence from a Pyrilidae species comprising a sequence encoding an Ecdysone receptor;
ii) a nucleotide sequence comprising the sequence set forth in SEQ ID NO: 1;
iii) a nucleotide sequence encoding a polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 2;
iv) a nucleotide sequence that hybridizes under stringent conditions to a sequence of i), ii), or iii);

b) further stably incorporating into the genome of said plant a second expression cassette,
wherein said expression cassette comprises a transcriptional regulatory region operably linked to a nucleotide sequence encoding said protein of interest, and
wherein said transcriptional regulatory region is activated by the ligand-receptor complex;
c) contacting said plant with a ligand which complexes with said receptor,
wherein said receptor-ligand complex interacts with the transcription regulatory region and induces gene expression of the protein of interest.

Claim 23
An expression vector comprising a promoter operably linked to a nucleotide sequence encoding an Ecdysone receptor wherein said nucleotide sequence encoding the Ecdysone receptor is selected from the group consisting of:

a) a nucleotide sequence from a Pyrilidae species comprising a sequence encoding an Ecdysone receptor;
b) a nucleotide sequence comprising the sequence set forth in SEQ ID NO: 1;
c) a nucleotide sequence encoding a polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 2;
d) a nucleotide sequence that hybridizes under stringent conditions to a sequence of a), b), or c).

Claim 25
An expression vector comprising a promoter operably linked to a nucleotide sequence encoding ultraspiracle, and said nucleotide sequence encoding ultraspiracle is selected from the group consisting of:

a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO: 3;
b) a nucleotide sequence encoding a polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 4;
c) a nucleotide sequence that hybridizes under stringent conditions to a sequence of a) or b).

Claim 1
A vector comprising a selection gene, yeast origin of replication or autonomously replicating sequence and DNA encoding a eukaryotic polypeptide other than yeast chelatin, said DNA encoding a eukaryotic polypeptide that is under the control of:

a) a yeast chelatin promoter,
b) a yeast chelatin transcription control sequence, or
c) a metal ion regulatory region of the yeast chelatin transcription control sequence, which is free of the yeast chelatin promoter,

whereby the vector is replicable in a suitable host.

Claim 14
An isolated DNA sequence comprising a yeast chelatin metal ion regulatory region free of the yeast chelatin promoter and free of DNA encoding yeast che latin.

Claim 1
A process for subjecting the transcription of a selected DNA sequence to external control under given environmental conditions which comprises the steps of:

A) providing a selected isolated structural gene that is transcriptionally responsive to a mouse metallothionein-I promoter/regulator DNA sequence under the given environmental conditions; and
B) operatively fusing the selected structural gene with said promoter/regulator DNA sequence.

Claim 2
In the genetic engineering process for securing transcription and expression of a selected isolated structural gene sequence in a mammalian host cell wherein said selected structural gene is stably incorporated as a chromosomal or extrachromosomal constituent of the host, the improvement comprising the step of:

operatively fusing with said selected structural gene sequence a mouse metallothionein-I promoter/regulator DNA sequence, which is responsive to environmental variations within the host cell in the concentration of ions of metal.

Claim 3
A fusion gene product, suitable for use in genetic transformation of a mammalian host cell, said product comprising:

a non mouse metallothionein structural gene sequence to be incorporated in said host cell operatively fused with a mouse metallothionein-I promoter/regulator DNA sequence.

Claim 1
A DNA sequence of less than 500 base pairs, said DNA sequence comprising the human MT-II transcriptional regulatory system further comprising the transcription initiation sequence.

Claim 5
A DNA construct capable of regulated expression of an inserted gene in a mammalian host, said construct comprising an extrachromosomal replication system recognized by a mammalian host, which replication system has been joined to a human MT-II gene inducible regulatory system in vitro, said construct having DNA sequence(s) defining at least one restriction site within the transcriptional control of the regulatory region for insertion of the gene.

Claim 10
A DNA construct useful for expression of a structural gene to produce a polypeptide in a mammalian host, said vector comprising (a) a replicon from bovine papilloma virus and (b) a regulatory system from a human MT-II gene comprising a promoter and a terminator and having at least one restriction site downstream from the promoter in the direction of transcription.

Claim 1
A nucleic acid promoter fragment isolated from the 5' flanking region upstream of the coding region of a cucumber chitinase/lysozyme gene that is inducible by application of benzo-1,2,3-thiadiazoles.

Claim 1
A chemically inducible nucleic acid promoter fragment isolated from the 5' flanking region upstream of the coding region of a tobacco PR-1a gene, wherein said promoter fragment comprises a nucleotide fragment of at least 603-bp adjacent to the coding region of said tobacco PR-1a gene, wherein said promoter fragment is inducible by application of a benzo-1,2,3-thiadiazole, an isonicotinic acid compound, or a salicylic acid compound.

Claim 3
A chemically inducible nucleic acid promoter fragment isolated from the 5' flanking region upstream of the coding region of an Arabidopsis PR-1 gene, wherein the coding region of said Arabidopsis PR-1 gene comprises the DNA sequence set forth in SEQ ID NO:33 or a DNA sequence which would encode the protein encoded by SEQ ID NO:33,
wherein said promoter fragment is inducible by application of a ben zo-1,2,3-thiadiazole, an isonicotinic acid compound, or a salicylic acid compound.

Claim 1
A method of inducing gene transcription in a plant or plant tissue, comprising the steps of:

A) transforming said plant or plant tissue, each with a chimeric gene comprising:
(i) a chemically inducible nucleic acid promoter fragment of at least 603-bp isolated from the 5' flanking region adjacent the coding region of a tobacco PR-1a gene, and
(ii) a coding sequence of interest operatively linked to said promoter fragment; and
B) exposing said transgenic plant or plant tissue to a benzo-1,2,3-thiadiazole, an isonicotinic acid compound, or a salicylic acid compound, whereby transcription of said coding sequence of interest is induced in said plant or plant tissue.

Claim 8
A method of inducing gene transcription in a plant or plant tissue, comprising the steps of:

A) transforming said plant or plant tissue, each with a chimeric gene comprising:
(i) a chemically inducible nucleic acid promoter fragment isolated from the 5' flanking region adjacent the coding region of an Arabidopsis PR-1 gene, wherein said Arabidopsis PR-1 gene comprises a DNA sequence that specifically hybridizes to SEQ ID NO:33 or wherein said Arabidopsis PR-1 gene comprises a DNA sequence that encodes the protein encoded by SEQ ID NO:33, and
(ii) a coding sequence of interest operatively linked to said promoter fragment; and
B) exposing said transgenic plant or plant tissue to a benzo-1,2,3-thiadiazole, an isonicotinic acid compound, or a salicylic acid compound, whereby transcription of said coding sequence of interest is induced in said plant or plant tissue.

Claim 1
An isolated DNA molecule comprising a nucleotide sequence selected from the following group:

a) a full-length chemically inducible promoter fragment comprising nucleotides 1 through 4258 of SEQ ID NO: 1;
b) an 815-bp long chemically inducible promoter fragment comprising nucleotides 3444 through 4258 of SEQ ID NO: 1; and
c) a 698-bp long chemically inducible promoter fragment comprising nucleotides 3561 through 4258 of SEQ ID NO: 1.

Claim 14
An isolated DNA molecule involved in inducibility of a chemically inducible promoter selected from the following group:

a) LS4 comprising nucleotides 3584 through 3593 of SEQ ID NO: 1;
b) LS7 comprising nucleotides 3614 through 3623 of SEQ ID NO: 1;
c) LS 10 comprising nucleotides 3644 through 3653 of SEQ ID NO: 1; and
d) a region spanning LS7-LS 1 0 and comprising nucleotides 3614 through 3653 of SEQ ID NO: 1.

Claim 1
An isolated promoter comprising a nucleotide sequence that initiates transcription in a plant cell, wherein said nucleotide sequence is selected from the group consisting of:

a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO: 3 or 4; and
b) a nucleotide sequence comprising the plant promoter sequence deposited in the plasmid designated as ATCC Accession No. 207139 or 207131.

Claim 5
A method for driving expression of a heterologous nucleotide sequence in a plant, said method comprising the steps of:

a) transforming a plant cell with an expression cassette comprising the heterologous nucleotide sequence operably linked to a promoter that initiates transcription in a plant cell, wherein said promoter is selected from the group consisting of:
i) a nucleotide sequence comprising the sequence set forth in SEQ ID NO: 3 or 4; and
ii) a nucleotide sequence comprising the plant promoter sequence deposited in the plasmid designated as ATCC Accession No. 207139 or 207131; and
b) regenerating a stably transformed plant from said plant cell.

Claim 9
A plant cell transformed with a DNA construct comprising a heterologous nucleotide sequence operably linked to a promoter that initiates transcription in said plant cell, wherein said promoter comprises a nucleotide sequence selected from the group consisting of:

a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO: 3 or 4; and
b) a nucleotide sequence comprising the plant promoter sequence deposited in the plasmid designated as ATCC Accession No. 207139 or 207131.

Claim 13
A plant stably transformed with a DNA construct comprising a heterologous nucleotide sequence operably linked to a promoter that initiates transcription in a plant cell, wherein said promoter comprises a nucleotide sequence selected from the group consisting of:

a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO: 3 or 4; and
b) a nucleotide sequence comprising the plant promoter sequence deposited in the plasmid designated as ATCC Accession No. 207139 or 207131.

Claim 1
An isolated nucleic acid molecule having a nucleotide sequence for a promoter that is capable of initiating transcription in a plant cell,
wherein said nucleotide sequence is selected from the group consisting of:

a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO: 1, 2, 3, 4, or 5;
b) a nucleotide sequence selected from the group consisting of sequences deposited as ATCC Accession No. _
c) a nucleotide sequence comprising at least 40 contiguous 10 nucleotides of the sequence set forth in SEQ ID NO: 1, 2, 3, 4, or 5; and
d) a nucleotide sequence that hybridizes under stringent conditions to a sequence of a), b), or c).

Claim 5
A method for inducing expression of a heterologous nucleotide sequence in a plant, said method comprising:

A) transforming a plant cell with a DNA construct comprising said heterologous nucleotide sequence operably linked to a promoter that is capable of initiating transcription in a plant cell in response to a stimulus,
B) regenerating a stably transformed plant from said plant cell, and
C) exposing said plant to said stimulus, wherein said promoter comprises a nucleotide sequence selected from the group consisting of:

a) a nucleotide sequence comprising the sequence set forth in 30 SEQ ID NO: 1, 2, 3, or 4;
b) a nucleotide sequence selected from the group consisting of sequences deposited as ATCC Accession No._
c) a nucleotide sequence comprising at least 40 contiguous 5 nucleotides of the sequence set forth in SEQ ID NO: 1, 2, 3, or 4; and
d) a nucleotide sequence that hybridizes under stringent conditions to a sequence of a), b), or c).

Claim 9
A method for constitutively expressing a heterologous nucleotide sequence in a plant, said method comprising:

A) transforming a plant cell with a DNA construct comprising said heterologous nucleotide sequence operably linked to a promoter that is capable of initiating constitutive transcription in a plant cell and
B) regenerating a stably transformed plant from said plant cell,
wherein said promoter comprises a nucleotide sequence selected from the group consisting of:

a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO: 5;
b) a nucleotide sequence selected from the group consisting of sequences deposited as ATCC Accession No._
c) a nucleotide sequence comprising at least 40 contiguous nucleotides of the sequence set forth in SEQ ID NO: 5; and
d) a nucleotide sequence that hybridizes under stringent conditions to a sequence of a), b), or c).

Claim 13
A plant cell stably transformed with a DNA construct comprising a heterologous nucleotide sequence operably linked to a promoter that is capable of initiating transcription in said plant cell,
wherein said promoter comprises a nucleotide sequence selected from the group consisting of :

a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO: 1, 2, 3, 4, or 5;
b) a nucleotide sequence selected from the group consisting of sequences deposited as ATCC Accession No._
c) a nucleotide sequence comprising at least 40 contiguous nucleotides of the sequence set forth in SEQ ID NO: 1, 2, 3, 4, or 5; and
d) a nucleotide sequence that hybridizes under stringent conditions to a sequence of a), b), or c).

Claim 17
A plant stably transformed with a DNA construct comprising a heterologous nucleotide sequence operably linked to a promoter that is capable of initiating transcription in a plant cell,
wherein said promoter comprises a nucleotide sequence selected from the group consisting of:

a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO: 1, 2, 3, 4, or 5;
b) a nucleotide sequence selected from the group consisting of sequences deposited as ATCC Accession No. _
c) a nucleotide sequence comprising at least 40 contiguous nucleotides of the sequence set forth in SEQ ID NO: 1, 2, 3, 4, or 5; and
d) a nucleotide sequence that hybridizes under stringent conditions to a sequence of a), b), or c).

Claim 22
An isolated nucleic acid molecule having a nucleotide sequence selected from the group consisting of:

a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO: 6, 8, 10, or 14;
b) a nucleotide sequence selected from the group consisting of sequences deposited as ATCC Accession No._
c) a nucleotide sequence encoding a polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 7, 9, 11, or 15;
d) a nucleotide sequence comprising at least 16 contiguous nucleotides of a sequence of a), b), or c); and
e) a nucleotide sequence that hybridizes under stringent conditions to a sequence of a), b), c), or d).

Claim 26
A method for creating or enhancing disease resistance in a plant, said method comprising:

A) transforming said plant with a DNA construct comprising a PR-1 sequence operably linked to a promoter that drives expression of a coding sequence in a plant cell and
B) regenerating stably transformed plants,
wherein said PR-1 sequence is selected from the group consisting of:

a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO: 6, 8, 10, or 14;
b) a nucleotide sequence selected from the group consisting of sequences deposited as ATCC Accession No._
c) a nucleotide sequence encoding a polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 7, 9, 11, or 15;
d) a nucleotide sequence comprising at least 16 contiguous nucleotides of a sequence of a), b), or c); and
e) a nucleotide sequence that hybridizes under stringent conditions to a se quence of a), b), c), or d).

Claim 33
A plant cell stably transformed with a DNA construct comprising a PR-1 sequence operably linked to a promoter that drives expression of a coding sequence in a plant cell, wherein said PR-1 sequence is selected from the group consisting of:

a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO: 6, 8, 10, or 14;
b) a nucleotide sequence selected from the group consisting of sequences deposited as ATCC Accession No._
c) a nucleotide sequence encoding a polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 7, 9, 11, or 15;
d) a nucleotide sequence comprising at least 16 contiguous nucleotides of a sequence of a), b), or c); and
e) a nucleotide sequence that hybridizes under stringent conditions to a sequence of a), b), c), or d).

Claim 34
A plant stably transformed with a DNA construct comprising a PR-1 sequence operably linked to a promoter that drives expression of a coding sequence in a plant cell, wherein said PR-1 sequence is selected from the group consisting of:

a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO: 6, 8, 10, or 14;
b) a nucleotide sequence selected from the group consisting of sequences deposited as ATCC Accession No._
c) a nucleotide sequence encoding a polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 7, 9, 11, or 15;
d) a nucleotide sequence comprising at least 16 contiguous nucleotides of a sequence of a), b), or c); and
e) a nucleotide sequence that hybridizes under stringent conditions to a sequence of a), b), c), or d).

Claim 1
A recombinant heat shock gene comprising a plant heat shock promoter and a heterologous structural gene whose expression is controllable thereby,
wherein said promoter comprises the consensus nucleotide sequence 5'-C-T-X-G-A-A-X-X-T-A-C-X-X-X-3',
where X is A,T,C or G.

Claim 2
A recombinant heat shock gene comprising a plant heat shock promoter and a heterologous structural gene whose expression is controlled thereby,
wherein said promoter comprises the consensus nucleotide sequence 5'-C-T-S-G-A-A-M-R-T-A-C-W-M-K-3',
where S is C or G; M is A or C; R is A or G; W is A or T; and K is T or G.

Claim 8
A vector comprising a fragment of DNA capable of functioning as a plant heat shock promoter wherein said fragment of DNA comprises the consensus nucleotide sequence 5'-C-T-X-G-A-A-X-X-T-A-C-X-X-X-3',
where X is A, T, C or G.

Claim 9
A vector comprising a fragment of DNA capable of functioning as a plant heat shock promoter wherein said fragment comprises the consensus nucleotide sequence of 5'-C-T-S-G-A-A-M-R-T-A-C-W-M-K-3',
where S is C or G; M is A or C; R is A or G; W is A or T; and K is T or G.

Claim 19
A bacterial strain containing therein recombinant DNA comprising:

a) a fragment of DNA capable of controlling heat shock expression of a gene in a plant; and
b) a structural gene oriented with respect to said fragment of DNA so as to be expressible under the control thereof
wherein said fragment of DNA capable of controlling heat shock expression of a gene in a plant comprises the consensus nucleotide sequence 5'-C-T-X-G-A-A-X-X-T-A-C-X-X-X-3',
where X is A, T, C or G.

Claim 20
A bacterial strain containing therein recombinant DNA comprising:

a) a fragment of DNA capable of controlling heat shock expression of a gene in a plant; and
b) a structural gene oriented with respect to said fragment of DNA so as to be expressible under the control thereof
wherein the fragment of DNA capable of controlling heat shock expression of a gene in a plant comprises the consensus nucleotide sequence 5'-C-T-S-G-A-A-M-R-T-A-C-W-M-K-3',
where S is C or G; M is A or C; R is A or G; W is A or T; and K is T or G.

Claim 34
A method for recognizing a plant cell containing recombinant DNA comprising the steps of:

1) transferring into said plant cell recombinant DNA comprising:
a) a heat shock promoter which comprises the consensus sequence 5'-C-T-X-G-A-A-X-X-T-A-C-X-X-X-3', where X is A, T, C, or G;
b) a transformation recognition gene under the control of said heat shock promoter capable of causing an observable or detectable reaction when expressed;
2) applying a stress to said plant cell in which it is desired to recognize those containing recombinant DNA, which stress is capable of inducing a response in said heat shock promoter; and
3) observing or detecting the reaction caused by expression of said transformation recognition gene under the control of said heat shock promoter to recognize said plant cell containing recombinant DNA including said tra nsformation recognition gene.

Claim 36
A method for recognizing a plant cell containing recombinant DNA comprising the steps of:

1) transferring into said plant cell recombinant DNA comprising:
a) a heat shock promoter which comprises the consensus sequence 5'-C-T-S-G-A-A-M-R-T-A-C-W-M-K-3',
where S is C or G; M is A or C; R is A or G; W is A or T; and K is T or G;
b) a transformation recognition gene under the control of said heat shock promoter capable of causing an observable or detectable reaction when expressed;
2) applying a stress to said plant cell in which it is desired to recognize those containing recombinant DNA, which stress is capable of inducing a response in said heat shock promoter; and
3) observing or detecting the reaction caused by expression of said transformation recognition gene under the control of said heat shock promoter to recognize said plant cell containing recombinant DNA including said transformation recognition gene.

Claim 1
A recombinant DNA plasmid comprising:

a) a vector,
b) a T-DNA fragment of a Ti-plasmid from an Agrobacterium strain, which T-DNA fragment is functional for insertion into a plant genome, and
c) a fragment of plant DNA which is capable of controlling gene expression in response to heat shock, and which fragment is inserted into said T-DNA fragment such that said T-DNA fragment retains functionality for insertion into a plant genome,
wherein said fragment of plant DNA comprises a plant heat shock promoter comprising the consensus nucleotide sequence 5'-C-T-X-G-A-A-X-X-T- A-C-X-X-X-3', wherein X is A, T, C or G.

Claim 17
A method for modifying a plant genome so as to allow expression of a structural gene under control of a soybean heat shock gene promoter fragment, comprising the steps of:

a) isolating a soybean heat shock gene promoter fragment comprising the consensus nucleotide sequence 5'-C-T-X-G-A-A-X-X-T-A-C-X-X-X-3', wherein X is A, T, C or G,
b) cloning said soybean heat shock gene promoter fragment into a T-DNA shuttle vector producing a recombinant DNA plasmid,
c) isolating a DNA fragment carrying foreign structural genes or soybean genes not naturally expressed under the regulatory control of a soybean heat shock gene promoter, and inserting said DNA fragment into said recombinant DNA plasmid at a position on the X-side of said soybean heat shock gene promoter producing a heat shock expression plasmid,
wherein said DNA fragment is oriented with respect to said soybean heat shock gene promoter as to be expressible under control thereof,
d) transforming said heat shock expression plasmid into a first bacterial strain capable of supporting replication of said heat shock expression plasmid,
e) mixing said bacterial strain capable of supporting replication of said heat shock expression plasmid with a second bacterial strain carrying a helper plasmid capable of transporting said heat shock expression plasmid into an Agrobacterium strain incapable of supporting replication of said heat shock expression plasmid, said Agrobacterium strain carrying a resident plasmid,
f) selecting for recombination between said heat shock expression plasmid and said resident plasmid giving a recombinant resident plasmid,
g) infecting a plant or a plant cell culture with said Agrobacterium strain containing and replicating said recombinant resident plasmid, and
h) selecting a plant or a plant cell culture comprising plant cells containing said foreign structural genes or said soybean genes under control of said soybean heat shock gene promoter transferred from said recombinant resident plasmid to said plant cells,
said foreign structural genes or said soybean genes being expressed following heat shock treatment or other stress treatment.

Claims 1, 8, and 19 are worded exactly the same as claims 1, 8 and 19 of the United States patent.

Claim 34
A method for obtaining a transient increase in expression level of a heterologous structural gene in a plant comprising:

1) inserting into said plant genome a DNA fragment comprising a promoter capable of inducing a heat shock response in a plant exposed to stress,
which promoter comprises the consensus nucleotide sequence 5'-C-T-X-G-A-A-X-X-T-A-C-X-X-X-5', where X is A, T, C or G,
said promoter being combined with said structural gene such that said structural gene is expressed under the control of said promoter, followed by 2) exposing said plant to stress such that said promoter responds thereby obtaining a transient increase in expression level of said structural gene in said plant.

Claim 50
A method for controlling the expression of a structural gene in a plant comprising the steps of:

1) transforming said plant with a DNA fragment comprising a DNA promoter sequence capable of controlling heat shock expression in a plant and a structural gene, wherein said DNA promoter sequence comprises the consensus nucleotide sequence 5'-C-T-X-G-A-A-X-X-T-A-C-X-X-X-3',
where X is A, T, C or G and said structural gene is oriented with respect to said DNA promoter sequence so as to be controllable thereby;
2) applying stress to said plant so as to activate said DNA promoter sequence and thereby cause the expression of said structural gene.

Claim 54
It is worded exactly the same as claim 34 of the United States patent.

Claim 1
A method for the spatial and temporal control of the expression of a gene of interest within a preselected discrete region of a cell mass or multicellular organism, comprising selectively heating a preselected discrete region of a cell mass or multicellular organism that includes cells that contain a genetically engineered gene of interest operably linked to a heat-inducible promoter, thereby inducing the expression of said gene of interest in the cells that are selectively heated.

Claim 9
A method of providing a therapeutic protein to selected cells in an animal, comprising the steps of:

A) introducing into cells of an animal a DNA molecule having a heat shock promoter sequence operably linked to and exerting regulatory control over a sequence encoding a therapeutic protein, and
B) activating said heat shock promoter sequence through the application of a focused ultrasound so that said DNA segment expresses a therapeutically effective amount of said therapeutic protein.

* Independent claims as filed of the European patent application EP 922110 A2 are the same as the independent, granted claims of the Australian patent.

Claim 1
A method for high level production of a polypeptide or protein in a transformed host cell comprising:

a) transforming a host cell with a structural gene encoding for a polypeptide or protein, under the control of an inducible heat shock promoter;
b) amplifying the copy number of said structural gene in said transformed host cells using an amplification system under the control of a promoter other than an inducible heat shock promoter;
c) inducing said inducible promoter by heat shock to said transformed host cells at a temperature and for a time sufficient to transcribe said structural gene;
d) allowing said heat shocked cells to recover at a lower temperature than said heat shock temperature and for a time sufficient to translate said transcribed structural gene, producing said polypeptide or protein.

Claim 10
A method for increasing the production of a structural polypeptide or protein in a transformed mammalian cell comprising the steps of:

a) culturing a dihydrofolate reductase (DHFR) deficient mammalian cell transformed with
(i) a DHFR gene under the control of a constitutive promoter, and
(ii) a structural gene encoding for a polypeptide or protein under the control of an inducible heat shock promoter, in a culture medium containing methotrexate, for a time sufficient to permit growth of said transformed mammalian cells;
b) inducing said heat shock promoter at a temperature and for a time sufficient to transcribe said structural gene; and
c) allowing said heat shocked cells to recover from said heat shock at a lower temperature and for a time sufficient to translate said transcribed structural gene, producing said polypeptide or protein.

Claim 17
A host cell cotransformed with:

a) a gene amplification system under the control of a constitutive promoter, and
b) a structural gene encoding for a polypeptide or protein under control of an inducible heat shock promoter.

Claim 1
A vector which is characterized in containing each of the following elements:

(1) a promoter which shows its action in the host to be used;
(2) regulatory region for regulating the action of the promoter of (1); and
(3) a region which codes for the 5'-untranslated region derived from cold-shock protein gene mRNA or a region which codes for the region where substitution, deletion, insertion or addition of at least one base is applied to the said untranslated region and
which contains a base sequence as shown in SEQ ID NO:1 in the Sequence Listing.

Claim 10
An isolated promoter consisting of a base sequence as shown in SEQ ID NO: 5 in the Sequence Listing.

Claim 11
An isolated promoter containing a base sequence as shown in SEQ ID NO: 5 in the Sequence Listing and
consisting of a base sequence having 135 or less bases,
wherein the promoter does not contain the region which is ascribed to mRNA.

Claim 1
A vector which is characterized in containing each of the following elements:

(1) a promoter which shows its action in the host to be used;
(2) regulatory region for regulating the action of the promoter of (1); and
(3) a region which codes for the 5'-untranslated region derived from cold-shock protein gene mRNA or a region which codes for the region where substitution, deletion, insertion or addition of at least one base is applied to the said untranslated region.

Claim 14
A promoter containing a base sequence as shown in SEQ ID NO:5 in the Sequence Listing and
consisting of a base sequence having 135 or less bases.

Claim 1
A DNA sequence comprising a nucleotide sequence from the first to the 3546th nucleotide in the nucleotide sequence shown in SEQ ID NO:1.

Claim 2
A DNA sequence comprising a nucleotide sequence from the 2418th to the 3541st nucleotide in the nucleotide sequence shown in SEQ ID NO:1.

Claim 3
A DNA sequence comprising a nucleotide sequence from the first to the 4120th nucleotide in the nucleotide sequence shown in SEQ ID NO:2

Claim 1
A DNA sequence having a nucleotide sequence from first to 3546th nucleotide in the nucleotide sequence shown in SEQ ID. No. 1, or a part thereof having a cold- inducible promoter activity, or a DNA sequence having the same nucleotide sequence as said DNA sequences except that one or more nucleotides are deleted or substituted, or one or more nucleotides are inserted or added, which DNA sequence has a cold-inducible promoter activity.

Claim 3
A DNA sequence having a nucleotide sequence from 2418th to 3541st nucleotide in the nucleotide sequence shown in SEQ ID. No. 1, or a part thereof having a cold - inducible promoter activity, or a DNA sequence having the same nucleotide sequence as the said DNA sequences except that one or more nucleotides are deleted or substituted, or one or more nucleotides are inserted or added, which DNA sequence has a cold-inducible promoter activity.

Claim 4
A cold-inducible promoter sequence having a nucleotide sequence from 2418th to 3541st nucleotide in the nucleotide sequence shown in SEQ ID. No. 1, or a DNA sequence having the same nucleotide sequence as said DNA sequence except that one or more nucleotides are deleted or substituted, or one or more nucleotides are inserted 45 or added, which DNA sequence has a cold-inducible promoter activity.

Claim 5
A DNA sequence having a nucleotide sequence from first to 4 120th nucleotide in the nucleotide sequence shown in SEQ ID. No. 2, or a part thereof having a cold- inducible promoter activity, or a DNA sequence having the same nucleotide sequence as the said DNA sequences except that one or more nucleotides are del eted or substituted, or one or more nucleotides are inserted or added, which DNA sequence has a cold-inducible promoter activity.

Claim 7
A probe comprising a DNA fragment having at least 18 consecutive nucleotides in the region from 45th to 839th nucleotide in the sequence shown in SEQ ID N0:3 in the Sequence Listing or a sequence complementary thereto.

*Independent claims as filed of the European application EP 812917 A1 are the same as the granted claims of the Australian patent. The only difference is in Claim 7 where instead of " having at least 18 consecutive nucleotide", the probe recited in the European application has "at least 15 consecutive nucleotides".

Claim 1
An isolated a-amylase promoter having cold-sensitive promoter activity, having a sequence comprising the 5.5 Kb EcoRI DNA fragment of Solanum tuberosum from the transformed E. coli strain, DH5alpha-gPAmy 351 (NCIMB Accession Number 40682).

Claim 12
An isolated a-amylase promoter having tuber-specific activity and having a sequence comprising SEQ ID NO: 1.

Claim 1
A promoter comprising a nucleotide sequence corresponding to the 5.5 Kb EcoR1 fragment isolated from Solanum tuberosum or a variant, homologue or fragment thereof.

Claim 2
A promoter comprising a nucleotide sequence corresponding to the 5.5 b EcoR1 fragment isolated from Solanum tuberosum or a variant, homologue or fragment thereof, but wherein at least a part of the promoter is inactivated.

Claim 3
A promoter comprising at least the nucleotide sequence shown as Seq.I.D. No. 1 or a variant, homologue or fragment thereof.

Claim 4
A promoter comprising the nucleotide sequence of any of one of the sequences shown as Seq.I.D.Nos. 4 - 17 or a variant, homologue or fragment thereof.

Claim 5
A promoter comprising a nucleotide sequence corresponding to the 5.5 Kb EcoR1 fragment isolated from Solanum tuberosum or a variant, homologue or fragment thereof, but wherein at least the nucleotide sequence shown as Seq.I.D. No. 1 is inactivated.

Claim 6
A promoter comprising a nucleotide sequence corresponding to the 5.5 Kb EcoR1fragment isolated from Solanum tuberosum or a variant, homologue or fragment thereof, but wherein at least any of one of the sequences shown as Seq.I.D.Nos. 2-16 is inactivated.

Claim 1
An isolated DNA molecule comprising a 5' regulatory region of a low temperature-responsive gene BN115 from Brassica napus, comprising nucleotides 961-1210 of SEQ ID NO:1.

Claim 7
An isolated cold inducible promoter, or a functional fragment thereof capable of regulating the expression of a gene in response to a change in temperature,
said cold inducible promoter comprising nucleotides 1-1271 of SEQ ID NO:1.

Claim 8
An isolated DNA molecule comprising a sequence of at least 15 contiguous nucleotides of a 5' regulatory region of a low temperature responsive gene BN115 from Brassica napus, as defined by nucleotides 1-1271 of SEQ ID NO:1.

Claim 13
An isolated enhancer, or a functional fragment thereof each capable of mediating the expression of a gene under the control of a promoter and said enhancer or fragment,
said enhancer obtained from the regulatory region of BN115 from Brassica napus.

Claim 16
An isolated negative regulatory element capable of repressing gene expression at 22� C., obtained from the regulatory region of BN115 from Brassica napus and comprising nucleotides 461-623 of SEQ ID NO:1.

Claim 1
An isolated DNA molecule comprising a 5' regulatory region of a low temperature-responsive gene BN115 from Brassica napus, comprising nucleotides 156-1362 of SEQ ID NO:1.

Claim 5
An isolated DNA molecule comprising a 5' regulatory region of a low temperature-responsive gene BN115 from Brassica napus, comprising nucleotides 1-1271 of SEQ ID NO:1.

Claim 7
An isolated cold inducible promoter, or a functional fragment thereof capable of regulating the expression of a gene in response to a change in temperature,
said cold inducible promoter comprising nucleotides 1-1271 of SEQ ID NO:1.

Claim 8
An isolated DNA molecule comprising a sequence defined by nucleotides 1-1271 of SEQ ID NO:1, a fragment thereof, or a nucleotide sequence that hybridizes to said sequence defined by nucleotides 1-1271 of SEQ ID NO:1 under stringent hybridization conditions, wherein said DNA molecule, said fragment thereof, or said nucleotide sequence exhibit low temperature activity, said stringent hybridization conditions are selected from:

i) hybridization at 4X SSC at 42�C followed by washing in 0.1X SSC at 65�C for an hour; and
ii) hybridization at 4X SSC, 50% formamide at 42�C followed by washing in 0.1X SSC at 65�C for an hour.

Claim 13
An isolated enhancer comprising a sequence defined by nucleotides 156-623 of SEQ ID NO:1, a functional fragment thereof, or a nucleotide sequence that hybridizes to said sequence defined by nucleotides 156-623 of SEQ ID NO:1 under stringent hybridization conditions, wherein said enhancer, fragment or nucleotide sequence are each capable of mediating expression of a gene, said stringent hybridization conditions are selected from:

i) hybridization at 4X SSC at 42�C followed by washing in 0.1X SSC at 65�C for an hour; and
ii) hybridization at 4X SSC, 50% formamide at 42�C followed by washing in 0.1X SSC at 65�C for an hour.

Claim 16
An isolated negative regulatory element capable of repressing gene expression at 22� C., obtained from the regulatory region of BN115 from Brassica napus and comprising nucleotides 461-623 of SEQ ID NO:1.

Claim 1
A method for obtaining a plant having a modified phenotype, said method comprising;

Transforming a host plant cell with a DNA construct under genomic integration conditions, wherein said construct comprises as operably linked components in the direction of transcription, a promoter region obtainable from a gene, wherein transcription of said gene is light-inducible in a plant chloroplast containing tissue, a DNA sequence of interest other than the native coding sequence of said gene, and a transcription termination region, wherein said components are functional in a plant cell,

Whereby said DNA construct becomes integrated into a genome of said plant cell; regenerating a plant from said transformed plant cell, and growing said plant under conditions whereby said DNA sequence of interest is expressed and a plant having said modified phenotype is obtained.

Claim 2
A method for altering the phenotype of chloroplast containing A DNA sequence comprising a light-inducible promoter derived from a myxobacterium, and
a nucleotide sequence containing a restriction site located to enable a nucleotide sequence which is to be expressed to be placed under the control of the said promoter.

Claim 1
A recombinant DNA sequence containing a light-inducible promoter derived from a myxobacterium.

Claim 2
A DNA sequence comprising a light-inducible promoter derived from a myxobacterium, and
a nucleotide sequence containing a restriction site located to enable a nucleotide sequence which is to be expressed to be placed under the control of the said promoter.

Claim 8
A recombinant DNA sequence comprising a light-inducible promoter derived from a myxobacterium,
operatively linked to a DNA sequence coding for a polypeptide to be expressed,
the coding DNA sequence not being operatively linked to the promoter in nature.

Claim 11
A method of producing a polypeptide which comprises:

a) positioning a gene sequence coding for the polypeptide-under the op erative control of a light-inducible promoter derived from a myxobacterium to produce a recombinant DNA molecule;
b) transforming a competent bacterial host with the said recombinant DNA molecule; and
c) subjecting the host to light to induce the promoter whereby the gene is expressed.

Claim 1
A DNA fragment containing the sequence of SEQ ID NO: 1 as a core sequence,
whereby expression of a gene placed downstream of said DNA fragment is repressed in the presence of light.

Claim 4
A promoter containing the nucleotide sequence of SEQ ID NO: 1 as a core sequence,
whereby expression of a gene placed downstream of said promoter is promoted in the dark but repressed in the presence of light.

Note: Actual filed independent claims of patent applications EP 1 077 257 & CA 2328139 AA are the same as the granted independent claims of the New Zealander patent.

Claim 1
A method for the expression of a structural gene in a plant cell under conditions of darkness comprising the steps of:

a) transforming said plant cell with a recombinant molecule comprising:
a plant, dark- and light-active maize Cab promoter/regulatory system,
wherein said promoter/regulatory system is the promoter/regulatory system of Cab AB1084 or cross-hybridizes with the promoter/regulatory system of Cab AB1 084 under stringent conditions, and
a heterologous plant-expressible structural gene that is under the regulatory control of said plant Cab promoter/regulatory system in said plant cell, and
b) maintaining conditions of darkness for expression of said structural gene in said transformed plant cell.

Claim 8
A method for enhancing the level of expression obtained in the dark of a plant-expressible gene in a plant cell by exposing said plant cell to conditions of illumination comprising the steps of:

a) transforming said plant cell with a recombinant molecule comprising:
a plant, dark- and light-active maize Cab promoter/regulatory system,
wherein said promoter/regulatory system is the promoter/regulatory system of Cab AB1084 or cross-hybridizes with the promoter/regulatory system of Cab AB1084 under stringent conditions, and
a heterologous plant-expressible structural gene that is under the regulatory control of said plant Cab promoter/regulatory system in said plant cell, and
b) maintaining conditions of darkness for expression of said structural gene in said transformed plant cell, and
c) applying conditions of illumination to said transformed plant cell such that enhancement by a factor of about 3- to 6-fold in expression of said structural gene is obtained.

Claim 15
A plant grown from a plant cell,
wherein said plant cell is transformed with a recombinant molecule comprising a plant, dark- and light-active maize Cab promoter/regulatory system,
wherein said promoter/regulatory system is the promoter/regulatory system of Cab AB1084 or cross-hybridizes with the promoter/regulatory system of Cab AB1084 under stringent conditions, and
a heterologous plant-expressible structural gene that is under the regulatory control of said plant Cab promoter/regulatory system in said plant cell.

Claim 1
A recombinant DNA molecule comprising:

a) a dark- and light-active Cab promoter/regulatory system which cross-hybridizes with the Cab AB1084 promoter/regulatory system under stringent conditions, and wherein said promoter/regulatory system:

(i) functions to direct the expression of a structural gene under its control during conditions of darkness and
(ii) is stimulated to direct enhanced expression of said structural gene under light conditions, and

b) a heterologous plant-expressible structural gene wherein said structural gene is placed under the regulatory control of said plant promoter/regulatory system.