Methods for transforming and expression screening of filamentous fungal cells with a DNA library
Abstract
The present invention relates to methods for expression screening of filamentous fungal transformants, comprising: (a) isolating single colony transformants of a DNA library introduced into E. coli; (b) preparing DNA from each of the single colony E. coli transformants; (c) introducing a sample of each of the DNA preparations of step (b) into separate suspensions of protoplasts of a filamentous fungus to obtain transformants thereof, wherein each transformant contains one or more copies of an individual polynucleotide from the DNA library; (d) growing the individual filamentous fungal transformants of step (c) on selective growth medium, thereby permitting growth of the filamentous fungal transformants, while suppressing growth of untransformed filamentous fungi; and (e) measuring activity or a property of each polypeptide encoded by the individual polynucleotides. The present invention also relates to isolated polynucleotides encoding polypeptides of interest obtained by such methods, to nucleic acid constructs, expression vectors, and recombinant host cells comprising the isolated polynucleotides, and to methods of producing the polypeptides encoded by the isolated polynucleotides.
- Inventors:
- Issue Date:
- Research Org.:
- National Renewable Energy Laboratory (NREL), Golden, CO (United States)
- Sponsoring Org.:
- USDOE
- OSTI Identifier:
- 1183451
- Patent Number(s):
- 9045748
- Application Number:
- 11/140,150
- Assignee:
- Novozymes, Inc. (Davis, CA)
- Patent Classifications (CPCs):
-
C - CHEMISTRY C12 - BIOCHEMISTRY C12N - MICROORGANISMS OR ENZYMES
C - CHEMISTRY C40 - COMBINATORIAL TECHNOLOGY C40B - COMBINATORIAL CHEMISTRY
- DOE Contract Number:
- AC36-98GO10337
- Resource Type:
- Patent
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES
Citation Formats
Teter, Sarah, Lamsa, Michael, Cherry, Joel, and Ward, Connie. Methods for transforming and expression screening of filamentous fungal cells with a DNA library. United States: N. p., 2015.
Web.
Teter, Sarah, Lamsa, Michael, Cherry, Joel, & Ward, Connie. Methods for transforming and expression screening of filamentous fungal cells with a DNA library. United States.
Teter, Sarah, Lamsa, Michael, Cherry, Joel, and Ward, Connie. Tue .
"Methods for transforming and expression screening of filamentous fungal cells with a DNA library". United States. https://www.osti.gov/servlets/purl/1183451.
@article{osti_1183451,
title = {Methods for transforming and expression screening of filamentous fungal cells with a DNA library},
author = {Teter, Sarah and Lamsa, Michael and Cherry, Joel and Ward, Connie},
abstractNote = {The present invention relates to methods for expression screening of filamentous fungal transformants, comprising: (a) isolating single colony transformants of a DNA library introduced into E. coli; (b) preparing DNA from each of the single colony E. coli transformants; (c) introducing a sample of each of the DNA preparations of step (b) into separate suspensions of protoplasts of a filamentous fungus to obtain transformants thereof, wherein each transformant contains one or more copies of an individual polynucleotide from the DNA library; (d) growing the individual filamentous fungal transformants of step (c) on selective growth medium, thereby permitting growth of the filamentous fungal transformants, while suppressing growth of untransformed filamentous fungi; and (e) measuring activity or a property of each polypeptide encoded by the individual polynucleotides. The present invention also relates to isolated polynucleotides encoding polypeptides of interest obtained by such methods, to nucleic acid constructs, expression vectors, and recombinant host cells comprising the isolated polynucleotides, and to methods of producing the polypeptides encoded by the isolated polynucleotides.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {Tue Jun 02 00:00:00 EDT 2015},
month = {Tue Jun 02 00:00:00 EDT 2015}
}
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