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. 1985 Feb;21(2):191-200.

[Synthesis and cloning of DNA complementary to mRNA of the bovine mammary gland]

[Article in Russian]
S I Gorodetskiĭ, D R Kiarshulite, V N Ivanov, T V Kapelinskaia, A S Kaledin
  • PMID: 2859232

[Synthesis and cloning of DNA complementary to mRNA of the bovine mammary gland]

[Article in Russian]
S I Gorodetskiĭ et al. Genetika. 1985 Feb.

Abstract

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.

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