Patent EP0225078A2 successfully refreshed from Google Patents.
EP0225078A2
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EP
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Composite expression cassettes for fungal transformation
Assignee: Panlabs Inc USA
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Filing: 1986-11-11
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Granted: —
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Expires: 2006-11-11
Abstract
Abstract Novel hybrid DNA sequences useful for transforming filamentous fungi are provided. The hybrid DNA constructs include an expression cassette capable of controlling the expression of a structural gene encoding a selectable marker functional in fungi. The expression cassette can comprise a fungal promoter sequence and a fungal terminator sequence not natively associated with the selectable marker gene. After transformation into a fungal host, the hybrid DNA may be maintained as an extrachromosomal element or, alternatively, integrated into the fungal genome. A plasmid, pGAN, embodying a preferred construct of the present invention was deposited at the American Type Culture Collection on 19 November 1985, and designated Accession No.53331.
Inventors
- 👤 John A. Rambosek
- 👤 David A. Finkelstein
- 👤 Jeanette Leach
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-09 02:04
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/14; C12N9/24; C12N9/2402; C12N9/2405; C12N9/2408; C12N9/2411; C12N9/2428; C12N15/00; C12N15/09; C12N15/63; C12N15/79; C12N15/80; C12N15/87; C12N15/90
Claims Analysis
Total Claims: 14
Claim 1
Independent Claim
57 words
A DNA construct comprising an expression cassette controlling the expression of a structural gene encoding a polypeptide capable of functioning as a selectable marker in a filamentous fungus, said expression cassette comprising a fungal promoter sequence not natively associated with said structural gene, wherein said hybrid DNA composition is capable of stable maintenance in said filamentous fungus.
Claim 2
Dependent Claim
57 words
A DNA construct comprising an expression cassette controlling the expression of a structural gene encoding a polypeptide capable of functioning as a selectable marker in a filamentous fungus, said expression cassette comprising a fungal promoter sequence not natively associated with said structural gene, wherein said hybrid DNA composition is capable of stable maintenance in said filamentous fungus.
Claim 3
Dependent Claim
23 words
A DNA construct according to claim 1, wherein the entire sequence of said construct is not native to the species of said fungus.
Claim 4
Dependent Claim
15 words
A DNA construct according to claim 1, wherein said gene encodes for neomycin phosphotransferase II.
Claim 5
Dependent Claim
77 words
A filamentous fungal host containing a hybrid DNA construct comprising a gene encoding a selectable marker, said gene placed in the correct orientation for transcription under control of a fungal promoter sequence and said gene followed by a fungal terminator sequence, wherein said gene is not natively associated with at least one of the sequences and said construct is stably maintained in said host as an extrachromosomal element or by integration into the genome of said host.
Claim 6
Dependent Claim
77 words
A filamentous fungal host containing a hybrid DNA construct comprising a gene encoding a selectable marker, said gene placed in the correct orientation for transcription under control of a fungal promoter sequence and said gene followed by a fungal terminator sequence, wherein said gene is not natively associated with at least one of the sequences and said construct is stably maintained in said host as an extrachromosomal element or by integration into the genome of said host.
Claim 7
Independent Claim
97 words
A method for stably transforming a filamentous fungus comprising the steps of: forming a vector by inserting a structural gene encoding a selectable marker functional in said fungus into an expression cassette comprising a fungal promoter sequence and terminator sequence, both sequences capable of controlling the expression of the inserted structural gene, wherein said structural gene is not natively associated with at least one of said promoter or terminator sequences; transforming said fungus with said vector to form transformants; growing said transformants under conditions requiring, for survival, functional expression of said structural gene; and isolating surviving transformants.
Claim 8
Independent Claim
97 words
A method for stably transforming a filamentous fungus comprising the steps of: forming a vector by inserting a structural gene encoding a selectable marker functional in said fungus into an expression cassette comprising a fungal promoter sequence and terminator sequence, both sequences capable of controlling the expression of the inserted structural gene, wherein said structural gene is not natively associated with at least one of said promoter or terminator sequences; transforming said fungus with said vector to form transformants; growing said transformants under conditions requiring, for survival, functional expression of said structural gene; and isolating surviving transformants.
Claim 9
Independent Claim
112 words
A method of modifying the level of a polypeptide produced by a filamentous fungus, said method comprising the steps of: i) forming a vector comprising a first expression cassette capable of controlling expression of a selection gene encoding a selectable marker, said expression cassette comprises a fungal promoter sequence, wherein at least one of said sequences is not natively associated with said first structural gene; ii) joining to said first expression cassette a second expression cassette capable of controlling expression of a structural gene encoding said polypeptide; iii) transforming said fungus with the vector to form transformants; and iv) growing said transformants under conditions suitable for expression of said second structural gene.
Claim 10
Independent Claim
112 words
A method of modifying the level of a polypeptide produced by a filamentous fungus, said method comprising the steps of: i) forming a vector comprising a first expression cassette capable of controlling expression of a selection gene encoding a selectable marker, said expression cassette comprises a fungal promoter sequence, wherein at least one of said sequences is not natively associated with said first structural gene; ii) joining to said first expression cassette a second expression cassette capable of controlling expression of a structural gene encoding said polypeptide; iii) transforming said fungus with the vector to form transformants; and iv) growing said transformants under conditions suitable for expression of said second structural gene.
Claim 11
Dependent Claim
21 words
A method of claim 6, wherein said polypeptide is glucoamylase, which is naturally produced by a wild-type strain of said fungus.
Claim 12
Dependent Claim
39 words
A hybrid DNA vector including a first expression cassette comprising a neomycin phosphotransferase II gene under transcriptional initiation and termination control of fungal promotion and termination sequences, wherein said vector is capable of stable transformation into a filamentous fungus.
Claim 13
Dependent Claim
39 words
A hybrid DNA vector including a first expression cassette comprising a neomycin phosphotransferase II gene under transcriptional initiation and termination control of fungal promotion and termination sequences, wherein said vector is capable of stable transformation into a filamentous fungus.
Claim 14
Dependent Claim
25 words
A hybrid DNA vector according to claim 8, wherein said fungal promoter and terminator sequences are from a glucoamylase gene or a glutamate dehydrogenase gene.
References Cited (Backward Citations)
No citation records indexed.
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