Construction and characterization of a cDNA clone containing a portion of the bovine prolactin sequence.
Poly(A)-containing RNA from the bovine anterior pituitary has been used as a template for the enzymatic synthesis of double-stranded cDNA. The resulting double-stranded cDNA was inserted into the Pst I site of pBR322 with the oligo(dG)-oligo(dC) tailing technique and subsequently cloned in E. coli chi 1776. Clones containing sequences complementary to prolactin mRNA were identified by colony hybridization with partially purified prolactin cDNA. A 250 base pair sequence from one prolactin positive clone was extensively characterized and shown to contain the coding information for amino acids 119-192 of authentic bovine prolactin. The recombinant DNA from this clone was covalently attached to diazotized aminocellulose and used to purify prolactin mRNA from a mixture of mRNAs.
D000818:Animals; D002417:Cattle; D003001:Cloning, Molecular; D004277:DNA, Single-Stranded / Q000096:biosynthesis; D004926:Escherichia coli / Q000378:metabolism; D005796:Genes; D009693:Nucleic Acid Hybridization; D010903:Pituitary Gland, Anterior / Q000378:metabolism*; D010957:Plasmids; D011388:Prolactin / Q000096:biosynthesis*; D012333:RNA, Messenger / Q000378:metabolism*; D014170:Transformation, Genetic
D004277:DNA, Single-Stranded; D012333:RNA, Messenger; D011388:Prolactin