[Synthesis and cloning of DNA complementary to mRNA of the bovine mammary gland].
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.
Gorodetskiĭ
S I
SI
Kiarshulite
D R
DR
Ivanov
V N
VN
Kapelinskaia
T V
TV
Kaledin
A S
AS
D000818:Animals; D002364:Caseins / Q000235:genetics; D002417:Cattle / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods*; D004247:DNA / Q000096:biosynthesis*; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D005815:Genetic Code; D008321:Mammary Glands, Animal / Q000378:metabolism*; D009693:Nucleic Acid Hybridization; D010957:Plasmids; D011061:Poly A / Q000235:genetics; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics*; D011995:Recombination, Genetic
D002364:Caseins; D012333:RNA, Messenger; D011061:Poly A; D004247:DNA
Sintez i klonirovanie DNK, komplementarnykh mRNK molochnoĭ zhelezy korovy.