Synthesis and cloning of DNA complementary to mRNA of the bovine mammary gland
Gorodetskiĭ, S.I.; Kiarshulite, D.R.; Ivanov, V.N.; Kapelinskaia, T.V.; Kaledin, A.S.
Genetika 21(2): 191-200
1985
ISSN/ISBN: 0016-6758 PMID: 2859232 Document Number: 247989
Poly(A+)mRNA from bovine mammary glands was used to synthesize double-stranded cDNAs that were subsequently inserted into the plasmid vector pBR322 at the Pst1 site by means of oligo(dG)-oligo(dC) tailing. After transfection of Escherichia coli JC5183, recombinant plasmid library containing 5400 clones was screened by serial rounds of colony hybridization in situ to total [23P] poly(A+)mRNA and electrophoretically homogenious [32P]16SmRNA of mammary glands. Then hybrid selection of mRNA and subsequent in vitro translation of selected mRNAs were performed. In this manner, recombinant clones coding for alpha S1- beta-, kappa-casein were identified. cDNA clones range in size from 35% for beta-casein, 65% for alpha S1-casein to about 95% for kappa-casein, in comparison with their respective mRNAs.