Granted patent and patent application filed by
Unilever Patent Holdings B.V.
Unilever discloses in its applications a process for
producing a transformed mould with A. tumefaciens. In the method, an
A. tumefaciens vector containing at least one expressible gene is
introduced into the mould.
Moulds are defined by the inventors to include fungi from all five
subdivisions of the division Eumycota. The examples in the disclosures include
fungi from the genera Aspergillus, Fusarium,
Trichoderma, Neurospora and Colletotrichum. The first
three genera contain species important in large scale fermentation and
production of homologous and heterologous proteins. Other species within these
genera are fungal pathogens and fungi that serve as important model organisms
for basic research.
Specific Patent Information
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Patent Number
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Title, Independent Claims and Summary of Claims
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Assignee
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US
6255115
- Earliest priority - 7 April 1997
- Filed - 7 October 1999
- Granted - 3 July 2001
- Expected expiry - 7 October 2019
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Title - Agrobacterium-mediated transformation of
moulds, in particular those belonging to the genus Aspergillus
| Claim 1
A process for producing a transformed mould, comprising: A) inserting a DNA
fragment containing at least one expressible gene to be introduced into a mould
into a vector of Agrobacterium tumefaciens between the T-DNA borders
present in that vector;
B) introducing the vector containing the DNA
fragment between the T-DNA borders into an Agrobacterium tumefaciens
strain containing a vir region in its DNA;
C) inducing
vir genes to release T-DNA containing said DNA fragment from said
Agrobacterium tumefaciens, and incubating the Agrobacterium
tumefaciens strain with the mould to be transformed; and
D) selecting
the transformed mould from the untransformed mould depending on the
characteristics of the introduced DNA or its expression product, and optionally
culturing the transformed mould.
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The United States patent claims
- a method for producing a transgenic mould by inserting into it at least an
expressible gene carried in an A. tumefaciens vector. A.
tumefaciens containing the vector and the mould are co-cultivated and the
transformed mould is selected.
Note that although the title of the patent refers to the genus
Aspergillus, the independent claims are not limited to
this genus or any other genera of filamentous fungi.
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Unilever Patent
Holdings B.V.
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EP
973917 B1
- Earliest priority - 7 April 1997
- Filed - 24 March 1998
- Granted - 3 March 2004
- Expected expiry - 23 March 2018
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Title - Agrobacterium-mediated transformation of
moulds, in particular those belonging to the genus Aspergillus
| Claim 1
A process for producing a transformed mould, characterized in that:
1) a DNA fragment containing at least one expressible gene to be introduced
into a mould is first cloned into a vector of Agrobacterium tumefaciens
between the T-DNA borders present in that vector, wherein T-DNA borders
are 24 basepair imperfect direct repeats flanking the T-DNA;
2)
the vector containing the DNA fragment between the T-DNA borders is introduced
into an Agrobacterium tumefaciens strain containing a vir
region in its DNA;
3) release of T-DNA containing said DNA fragment from
said Agrobacterium tumefaciens by addition of a vir-inducing
compound, and the Agrobacterium tumefaciens strain is incubated with
the mould to be transformed; and
4) the transformed mould is selected from
the untransformed mould depending on the characteristics of the introduced DNA
or its expression product, and optionally the transformed mould is cultured.
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The claims as filed in the granted EP patent are directed to the same subject
matter as the granted claims of the United States patent.
Designated contracting States at the time of grant are: Switzerland, Germany,
Spain, France, United Kingdom, Italy, Liechtenstein.
* As the independent claims of the United States patent and the European
patent application are worded slightly different, the claims are presented
independently. However, the claims have practically the same scope.
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EP 973917 A1
- Earliest priority - 7 April 1997
- Filed - 24 March 1998
- Granted as EP 973917 B1 (see above)
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Title - Agrobacterium-mediated transformation of
moulds, in particular those belonging to the genus Aspergillus
| Claim 1 (granted with minor changes)
A process for producing a transformed mould, characterized in that:
1) a DNA fragment containing at least one expressible gene to be introduced
into a mould is first cloned into a vector of Agrobacterium tumefaciens
between the T-DNA borders present in that vector;
2) the vector containing
the DNA fragment between the T-DNA borders is introduced into an
Agrobacterium tumefaciens strain containing a vir region in
its DNA;
3) release of T-DNA containing said DNA fragment from said
Agrobacterium tumefaciens by addition of a vir-inducing
compound, and the Agrobacterium tumefaciens strain is incubated with
the mould to be transformed; and
4) the transformed mould is selected from
the untransformed mould depending on the characteristics of the introduced DNA
or its expression product, and optionally the transformed mould is cultured.
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Remarks
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- Related applications also filed in Brazil (BR 9807941 A),
Indonesia (ID 22929), and South Africa (ZA 9802905
A).
- National phase entry of WO 1998/45455 in Australia (AU
74283/98) has lapsed on 23 December 1999.
- National phase entry of WO 1998/45455 in Canada (CA
2286307) and Japan (JP 2001/518786 T2) are still
pending.
- National phase entry of WO 1998/45455 in China (CN
1151264) has been granted on 26 May 2004.
- Related patent family of
WO
1999/32641 'A process for site-directed integration of multiple copies of a
gene in a mould' describes a method of site-specific transformation of mould
using a rare restriction endonuclease system. The first independent claim
does not recite transformation using Agrobacterium.
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Note: Patent information on this page was last updated on 9 February 2006.
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time it was collated. New patents and patent applications, altered
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