doi: 10.1073/pnas.75.8.3727.
A bacterial clone synthesizing proinsulin
- PMID: 358198
- PMCID: PMC392859
- DOI: 10.1073/pnas.75.8.3727
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A bacterial clone synthesizing proinsulin
Proc Natl Acad Sci U S A.
1978 Aug.
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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