Open AccessCloning and nucleotide sequence of DNA coding for bovine preproparathyroid hormone.
Author(s) -
Henry M. Kronenberg,
Barbara E. McDevitt,
Joseph A. Majzoub,
Jeremy Nathans,
Phillip A. Sharp,
John T. Potts,
Alexander Rich
Publication year - 1979
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.76.10.4981
Subject(s) - coding region , nucleic acid sequence , recombinant dna , biology , nucleotide , microbiology and biotechnology , dna , plasmid , molecular cloning , genetics , peptide sequence , gene
We have cloned in Escherichia coli a DNA copy of mRNA coding for bovine preproparathyroid hormone. Double-stranded DNA was inserted into the Pst I site in plasmid pBR322 by using the poly(dG)-poly(dC) homopolymer extension technique to join the DNA molecules. Recombinant plasmids coding for preproparathyroid hormone were identified by the plasmid's ability to arrest specifically the translation of preproparathyroid hormone mRNA. The nucleotide sequence of the largest recombinant was determined by using both chemical and enzymatic techniques. The parathyroid insert contains 470 nucleotides--102 nucleotides from the 5' noncoding region of the mRNA, 345 nucleotides representing the entire coding region, and 23 nucleotides from the 3' noncoding region. The coding sequence clarifies the hormone's amino acid sequence, which has been disputed. Codon usage is discussed.