Method for identifying genes encoding secreted or membrane-associated proteins
US5952171A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | Nov 19, 1996 |
| Grant date | Sep 14, 1999 |
| Priority date | — |
| Expiry date | Nov 19, 2016 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12N2830/38
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
The invention features a method for identifying a cDNA nucleic acid encoding a mammalian protein having a signal sequence, which method includes the following steps: (a) providing library of mammalian cDNA; (b) ligating the library of mammalian cDNA to DNA encoding alkaline phosphatase lacking both a signal sequence and a membrane anchor sequence to form ligated DNA; (c) transforming bacterial cells with the ligated DNA to create a bacterial cell clone library; (d) isolating DNA comprising the mammalian cDNA from at least one clone in the bacterial cell clone library; (e) separately transfecting DNA isolated from clones in step (d) into mammalian cells which do not express alkaline phosphatase to create a mammalian cell clone library wherein each clone in the mammalian cell clone library corresponds to a clone in the bacterial cell clone library; (f) identifying a clone in the mammalian cell clone library which express alkaline phosphatase; (g) identifying the clone in the bacterial cell clone library corresponding to the clone in the mammalian cell clone library identified in step (f); and (h) isolating and sequencing a portion of the mammalian cDNA present in the bacterial cell l…
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