Method for identification and isolation of DNA encoding a desire

Details Method for identification and isolation of DNA encoding a desire Method for identification and isolation of DNA encoding a desire

1984-09-19

1987-06-23

Kepplinger, Esther M.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

435 68, 4351723, 435240, 435317, 935 78, 935 11, 935 56, C12Q 168, C12P 2100, C12N 1500, C12N 500

Patent

active

046752854

ABSTRACT:
A method for identifying and isolating clones containing DNA coding for a desired protein is described. DNA prepared from a cell that expresses the desired protein is inserted into an isolation expression vector having means for replication (as a means of producing DNA) and a suitable promoter for expression of said DNA in a predetermined mammalian host cell as well as means for replication in a bacterial cell. The transient expression vector is then inserted into a bacterial cell for replication of the DNA. Pools of DNA, prepared from a predetermined number of bacterial clones so that the nucleic acids (DNA and RNA) is substantially free of other bacterial contaminants are transfected or microinjected into mammalian host cells and conditioned medium from growing such cells is tested for the presence of the desired protein. Positive pools are selected and the clones used to make the pool are screened to identify and isolate the clone containing the desired DNA.

REFERENCES:
patent: 4446235 (1984-05-01), Seeburg
patent: 4517294 (1985-05-01), Bock et al.
patent: 4520106 (1985-05-01), Kado
Okayama et al., Mol. Cell Biol. vol. 3, No. 2, pp. 280-289 (1983).
Gray et al., Nature 295 pp. 503-508 (1982).
Yokoto et al., Proc. Natl. Sci. USA, vol. 81, pp. 1070-1074 (1984).

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