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Nucleic Acids Research, 1980, Vol. 8, No. 7 1561-1573
© 1980

Articles

Construction and characterization of a cDNA clone containing a portion of the bovine prolactin sequence

John H. Nilson+, Arlen R. Thomason*, Sarah Horowitz+, Nancy L. Sasavage+, John Blenis+, Robert Albers+,{dagger}, Winston Salser* and Fritz M. Rottman+

+Department of Biochemistry, Michigan State University East Lansing, MI 48824, USA *Department of Biology, University of California Los Angeles, CA 90024, USA

Received March 6, 1980.

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 ol igo(dG)-ol igo(dC) tailing technique and subsequently cloned In E. coli x1776. 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.

{dagger}Present address: Department of Chemistry, Calvin College, Grand Rapids, Michigan 49506


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