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PMID: 3511472 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Short synthetic oligodeoxyribonucleotide leader sequences enhance accumulation of human proinsulin synthesized in Escherichia coli.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 83 ·No. 3 ·1986-02-00 ·Pages 561-5

Sung WL, Yao FL, Zahab DM, Narang SA

Abstract

Enhanced accumulation of human proinsulin synthesized in Escherichia coli has been achieved by inserting a short leader of homooligopeptide at the amino end of proinsulin. Out of 20 amino acid oligomers studied, (Ala)6, (Asn)6, (Cys)7, (Gln)7, (His)6, (Ser)6, and (Thr)6 leaders were the most effective, with the yield of proinsulin ranging between 6% and 26% of the total bacterial protein. These constructions were made by inserting a synthetic oligodeoxyribonucleotide duplex, coding for a small homooligopeptide, between a synthetic proinsulin gene and an eight-codon beta-galactosidase gene residue in vector pUC8. Cyanogen bromide cleavage of the 102 amino acid fused polypeptide yielded a species identical to authentic proinsulin, as judged by NaDodSO4/PAGE and radioimmunoassay.

MeSH Terms

Escherichia coli/metabolism Genes Humans Oligodeoxyribonucleotides/metabolism Plasmids Proinsulin/biosynthesis,genetics Protein Sorting Signals/biosynthesis Recombinant Proteins/biosynthesis

Chemicals

Oligodeoxyribonucleotides Protein Sorting Signals Recombinant Proteins Proinsulin

Authors & Affiliations

4 authors, click to expand affiliations / ORCID

Sung W L

Yao F L

Zahab D M

Narang S A

References (16)

16 references, click to expand

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Article Info

Journal

Proceedings of the National Academy of Sciences of the United States of America

Abbr.

Proc Natl Acad Sci U S A

ISSN

0027-8424

DOI

10.1073/pnas.83.3.561

Published

1986-02-00

Pages

561-5

Language

English

Region

United States

NLM ID

7505876

PMCID

PMC322903

Subset

Databases

GENBANK

M12688

External Links

PubMed source DOI: 10.1073/pnas.83.3.561 PMC Full Text View journal details

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Short synthetic oligodeoxyribonucleotide leader sequences enhance accumulation of human proinsulin synthesized in Escherichia coli.

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