A bacterial clone synthesizing proinsulin.

1978/08/01 by Lydia Villa‐Komaroff, Argiris Efstratiadis, S Broome +5 · 4 citations
Medicine · Biochemistry, Genetics and Molecular Biology · #Pancreatic function and diabetes #Diabetes and associated disorders #Biochemical and Molecular Research

paper · doi:10.1073/pnas.75.8.3727

openalex publication_date 1978/08/01 · openalex created_date 2025/10/10 · openalex updated_date 2026/07/25

Abstract

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.

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