One-step gene amplification by Mu-mediated transposition of E. coli genes to a multicopy plasmid

Diego de Mendoza, David Clark, John E. Cronan

Research output: Contribution to journalArticlepeer-review

Abstract

A general in vivo method to amplify the number of copies of a specific gene in one step is described. The method is directly applicable to any selectable gene of Escherichia coli and is based on the Mu-mediated transposition of segments of host chromosomes into the conjugative, multicopy plasmid R6K. Using this method we have cloned the β-hydroxydecanoyl thioester dehydrase structural gene, fabA, into the R6K plasmid. Strains carrying the resultant plasmid produced 13 to 21 times more dehydrase than control strains.

Original languageEnglish (US)
Pages (from-to)27-32
Number of pages6
JournalGene
Volume15
Issue number1
DOIs
StatePublished - Oct 1981

Keywords

  • Molecular cloning
  • R6K vector
  • fabA
  • in vivo genetic engineering
  • β-hydroxydecanoyl thioester dehydrase

ASJC Scopus subject areas

  • Genetics

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