Synthesis of DNA coding for human proinsulin.

Published

Journal Article

A chemical-enzymatic synthesis of 271- and 286-bp DNA duplexes, each of which contains the entire sequence coding for human proinsulin has been accomplished. In addition to the coding sequence, the 271-bp fragment carries translation initiation and termination signals plus EcoRI-HindIII restriction enzyme sites for insertion into an appropriate plasmid vector. The 286-bp fragment also contains a Shine-Dalgarno (SD) sequence preceding an ATG codon. Employing the 286-bp polynucleotide, the 568-bp tandem proinsulin gene has been obtained. The synthesis of these DNA fragments involved preparation of 42 oligonucleotides by a rapid N-methylimidazolide phosphotriester method and enzymatic conversion of the oligonucleotides into the gene subfragments, which were cloned separately and fused to yield the desired DNAs coding for proinsulin. The proinsulin gene fragments were cloned in Escherichia coli and shown to have the correct sequences.

Full Text

Duke Authors

Cited Authors

  • Ovchinnikov, YA; Efimov, VA; Ivanova, IN; Reverdatto, SV; Skiba, NP; Chakhmakhcheva, OG

Published Date

  • November 1, 1984

Published In

Volume / Issue

  • 31 / 1-3

Start / End Page

  • 65 - 78

PubMed ID

  • 6098538

Pubmed Central ID

  • 6098538

International Standard Serial Number (ISSN)

  • 0378-1119

Digital Object Identifier (DOI)

  • 10.1016/0378-1119(84)90196-3

Language

  • eng

Conference Location

  • Netherlands