Construction of bacterial plasmids that contain the nucleotide sequence for bovine corticotropin-β-lipotropin precursor

S. Nakanishi, A. Inoue, T. Kita, S. Numa, A. C. Chang, S. N. Cohen, J. Nunberg, R. T. Schimke

Research output: Contribution to journalArticlepeer-review

Abstract

mRNA that encodes the common peptide precursor for the hormones corticotropin and β-lipotropin was purified from the neurointermediate lobe of bovine pituitaries, and double-stranded cDNA species synthesized from this template were cloned in Escherichia coli χ 1776 by inserting them into the Pst I endonuclease cleavage site of the pBR322 plasmid using poly(dG)poly(dC) homopolymeric extensions. Certain of the cloned cDNA inserts contain nucleotides corresponding to the complete amino acid sequence of bovine corticotropin and a coding sequence that corresponds to at least the first portion of bovine β-lipotropin. The nucleotide sequences coding for corticotropin and β-lipotropin are separated on the cDNA by a 6-base-pair sequence encoding lysine and arginine, indicating that the carboxyl terminus of corticotropin is connected on the precursor peptide with the amino terminus of β-lipotropin by these two amino acids. In addition, the cloned cDNA insert is characterized by an unusually high C+G nucleotide base content as well as by a number of DNA sequence duplications.

Original languageEnglish
Pages (from-to)6021-6025
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume75
Issue number12
DOIs
StatePublished - 1978

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