Open AccessA bacterial clone synthesizing proinsulin.
Author(s) -
Lydia VillaKomaroff,
Argiris Efstratiadis,
Stephanie Broome,
Peter T. Lomedico,
Richard Tizard,
Stephen P. Naber,
William L. Chick,
Walter Gilbert
Publication year - 1978
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.75.8.3727
Subject(s) - complementary dna , proinsulin , plasmid , biology , escherichia coli , microbiology and biotechnology , amino acid , peptide sequence , pbr322 , cdna library , biochemistry , dna , gene , insulin , endocrinology
We have cloned double-stranded cDNA copies of a rat preproinsulin messenger RNA in Escherichia coli chi1776, using the unique Pst endonuclease site of plasmid pBR322 that lies in the region encoding amino acids 181-182 of penicillinase. This site was reconstructed by inserting the cDNA with an oligo(dG)-oligo(dC) joining procedure. One of the clones expresses a fused protein bearing both insulin and penicillinase antigenic determinants. The DNA sequence of this plasmid shows that the insulin region is read in phase; a stretch of six glycine residues connects the alanine at position 182 of penicillinase to the fourth amino acid, glutamine, of rat proinsulin.