Synthesis of a human insulin gene V. Enzymatic assembly, cloning and characterization of the human proinsulin DNA

R. Brousseau, R. Scarpulla, W. Sung, H. M. Hsiung, S. A. Narang, Ray Wu

Research output: Contribution to journalArticlepeer-review

35 Scopus citations

Abstract

To form a 258-bp sequence coding for human proinsulin, 41 synthetic deoxyribo-oligonucleotide fragments of 11 to 15 nucleotides in length were assembled by enzymatic methods. The coding sequence is preceded by ATG and followed by TGA for translation start and stop signals, and terminated in an EcoRI and a BamHI recognition sequence. The complete synthetic sequence was ligated to a plasmid and cloned in Escherichia coli. The cloned DNA was shown to have the correct human proinsulin coding sequence.

Original languageEnglish (US)
Pages (from-to)279-289
Number of pages11
JournalGene
Volume17
Issue number3
DOIs
StatePublished - Mar 1982

Funding

We wish to thank Douglas H. Smith and J. Michniewicz for excellent technicaals sistance and Li-He Guo for valuable help with the sequence analysis of the cloned proinsulin gene. The work was supported by research grant AM21801 from the National Institute of Health, U.S. Public Health Service, from the National Research Council of Canada (NRCC NO 19599)a nd a fellowship grant DRG-323F (to R.C.S.) from the Damon Runyon-Walter Winchell Cancer Fund.

Keywords

  • Recombinant DNA
  • plasmid vectors
  • start and stop signals

ASJC Scopus subject areas

  • Genetics

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