Methods for transforming and expression screening of filamentous fungal cells with a DNA library
US9045748B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | May 26, 2005 |
| Grant date | Jun 2, 2015 |
| Priority date | — |
| Expiry date | Apr 25, 2026 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC40B40/02
- WIPO fieldBiotechnology
- WIPO sectorChemistry
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 polynucleotide…
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.