Recombinant primate granulocyte macrophage-colony stimulating fa

Details Recombinant primate granulocyte macrophage-colony stimulating fa Recombinant primate granulocyte macrophage-colony stimulating fa

1995-06-06

1999-08-24

Mertz, Prema

Drug, bio-affecting and body treating compositions

Lymphokine

530351, 536 235, 435 695, 435 712, 435325, 4352523, 4353201, 435471, A61K 3819, C07K 14535, C12N 1527

Patent

active

059422211

ABSTRACT:
A method for preparing and isolating a transformation vector containing CSF/cDNA is described. The method comprises: preparing RNA from a cell that produces CSF; preparing polyadenylated messenger RNA from said RNA; preparing single stranded cDNA from said messenger RNA; converting the single stranded cDNA to double stranded cDNA; inserting the double stranded cDNA into transformation vectors and transforming bacteria with said vector to form colonies; picking pools of 200 to 500 colonies each and isolating plasmid DNA from each pool; transfecting the plasmid DNA into suitable host cells for expressing CSF protein; culturing the transfected cells and assaying the supernatant for CSF activity; and selecting CSF positive pools and screening the colonies used to make the pool to identify a colony having CSF activity. Also described are a cDNA coding for a protein having CSF activity (i.e. CSF/cDNA), a microorganism or cell line transformed with a recombinant vector containing such CSF/cDNA, and a method for producing CSF protein by expressing said CSF/cDNA by culturing a microorganism or cell line.

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