SYNTHESIS AND CLONING OF DNA COMPLEMENTARY TO MESSENGER-RNA FROM BOVINE MAMMARY-GLANDS
- 1 January 1985
- journal article
- research article
- Vol. 21 (2) , 191-200
Abstract
Poly(A+)mRNA from bovine mammary glands was used to synthesize double-stranded c[complementary]DNA 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 mRNA were performed. In this manner, recombinant clones coding for .alpha.s1-, .beta.-, x-casein were identified. cDNA clones range in size from 35% for .beta.-casein, 65% for .alpha.s1-casein to about 95% for x-casein, in comparison with their respective mRNA.This publication has 12 references indexed in Scilit:
- Characterization and Cloning of the mRNAs Specific for the Lactating Mouse Mammary GlandEuropean Journal of Biochemistry, 1982
- Characterization of the translation products of the major mRNA species from rabbit lactating mammary glands and construction of bacterial recombinants containing casein and α-lactalbumin complementary DNABiochemical Journal, 1982
- The construction, identification and partial characterization of plasmids containing guinea-pig milk protein complementary DNA sequencesBiochemical Journal, 1981
- Construction and preliminary characterization of the rat casein and alpha-lactalbumin cDNA clones.Journal of Biological Chemistry, 1981
- Isolation and Characterization of mRNA from Mammary Gland of Lactating CowAgricultural and Biological Chemistry, 1981
- Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose.Proceedings of the National Academy of Sciences, 1980
- Studies on the intracellular segregation of polyribosome-associated messenger ribonucleic acid species in the lactating guinea-pig mammary glandBiochemical Journal, 1979
- A rapid sensitive assay for specific protein synthesis in cells and in cell-free translations: Use of Staphylococcus aureus as an adsorbent for immune complexesAnalytical Biochemistry, 1979
- A rapid procedure for isolation of large quantities of Escherichia coli DNA polymerase I utilizing a lambdapolA transducing phage.Journal of Biological Chemistry, 1979
- Isolation and characterization of purified rat casein messenger ribonucleic acidsBiochemistry, 1976