Cloning and amplification of the .beta.-glucosidase gene of Trichoderma reesei
First Claim
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1. A process for expressing extracellular β
- -glucosidase from DNA encoding an extracellular β
-glucoidase which DNA is obtained from a microorganism selected from the group consisting of Trichoderma, Aspergillus, and Neurospora, and wherein said DNA sequence or a portion thereof is capable of amplification by PCR with SEQ ID NO;
3 and SEQ ID NO;
4, wherein the amplification conditions are denaturation at 95°
C. for 10 minutes, annealing at 50°
C. for 2 minutes and extension at 65°
C. for 10 minutes for 30 cycles and wherein the amplification product is a DNA sequence that comprises a DNA sequence of about 700 base pairs from about position 471 to about position 1171 of SEQ ID NO;
1, comprising;
expressing said extracellular β
-glucosidase from a DNA insert through recombinant techniques in a filamentous fungus, wherein the inserted DNA comprises all of the coding region of a fungal β
-glucosidase gene and sequence necessary for the β
-glucosidase gene'"'"'s transcription and translation, and wherein said filamentous fungus is selected from the group consisting of Trichoderma, Aspergillus, Neurospora, Humicola and Penicillium.
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Abstract
A process for expressing extracellular β-glucosidase in a filamentous fungus by expressing a fungal DNA sequence encoding enhanced, deleted or altered β-glucosidase in a recombinant host microorganism is disclosed. Recombinant fungal cellulase compositions containing enhanced, deleted or altered expression of β-glucosidase is also disclosed.
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Citations
19 Claims
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1. A process for expressing extracellular β
- -glucosidase from DNA encoding an extracellular β
-glucoidase which DNA is obtained from a microorganism selected from the group consisting of Trichoderma, Aspergillus, and Neurospora, and wherein said DNA sequence or a portion thereof is capable of amplification by PCR with SEQ ID NO;
3 and SEQ ID NO;
4, wherein the amplification conditions are denaturation at 95°
C. for 10 minutes, annealing at 50°
C. for 2 minutes and extension at 65°
C. for 10 minutes for 30 cycles and wherein the amplification product is a DNA sequence that comprises a DNA sequence of about 700 base pairs from about position 471 to about position 1171 of SEQ ID NO;
1, comprising;expressing said extracellular β
-glucosidase from a DNA insert through recombinant techniques in a filamentous fungus, wherein the inserted DNA comprises all of the coding region of a fungal β
-glucosidase gene and sequence necessary for the β
-glucosidase gene'"'"'s transcription and translation, and wherein said filamentous fungus is selected from the group consisting of Trichoderma, Aspergillus, Neurospora, Humicola and Penicillium. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
- -glucosidase from DNA encoding an extracellular β
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11. A process for expressing extracellular β
- -glucosidase, comprising a) inserting DNA encoding an extracellular β
-glucosidase gene obtained from a microorganism selected from the group consisting of Trichoderma, Aspergillus, and Neurospora, wherein said DNA sequence or a portion thereof is capable of amplification by PCR with SEQ ID NO;
3 and SEQ ID NO;
4, wherein the amplification conditions are denaturation at 95°
C. for 10 minutes, annealing at 50°
C. for 2 minutes and extension at 65°
C. for 10 minutes for 30 cycles and wherein the amplification product is a DNA sequence that comprises a DNA sequence of about 700 base pairs from about position 471 to about position 1171 of SEQ ID NO;
1, and wherein said host fungus is selected from the group consisting of Trichoderma, Aspergillus, Neurospora, Humicola and Penicillium;
b) isolating transformants having enhanced β
-glucosidase expression as compared to a non-transformed filamentous host;
c) culturing said transformants under conditions to permit growth of said transformants; and
d) isolating a fungal cellulase composition containing the β
-glucosidase from said transformants. - View Dependent Claims (12, 13, 14)
- -glucosidase, comprising a) inserting DNA encoding an extracellular β
-
15. A probe for use in detecting nucleic acid sequences encoding β
- -glucosidase from a filamentous fungi comprising SEQ ID. NO;
1. - View Dependent Claims (16)
- -glucosidase from a filamentous fungi comprising SEQ ID. NO;
- 17. A polynucleotide sequence comprising the nucleic acid sequence of SEQ ID NO:
Specification