PCR based gene engineering of the Vibrio harveyi lux operon and the Escherichia coli trp operon provides for biochemically functional ... View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

1991-04

AUTHORS

Philip J. Hill, Simon Swift, Gordon S. A. B. Stewart

ABSTRACT

The polymerase chain reaction (PCR) was applied to clone the luxA and luxB genes from Vibrio harveyi, and the trp poL (promoter operator leader) region and the trpB and trpA genes from Escherichia coli. PCR-derived luxA/B and trpB/A genes were shown to express bacterial luciferase and tryptophan synthase respectively, when introduced into E. coli on a plasmid cloning vehicle. The trp poL was used successfully to control the expression of lac alpha, luxAB, trpB and trpA. PCR was also used to construct a functional luxAB translational fusion protein. Primers for this were designed to facilitate precise gene fusion and to provide a silent mutation within an EcoRI site in the luxB gene. Production of functional genes was verified in vitro and in vivo using polyacrylamide gel electrophoresis (PAGE) analysis of transcription-translation products and crude cell extracts, and by monitoring enzyme activity. More... »

PAGES

41-48

References to SciGraph publications

  • 1981-02. Attenuation in the control of expression of bacterial operons in NATURE
  • 1989-04. New Clues on Folding in NATURE BIOTECHNOLOGY
  • Journal

    TITLE

    Molecular Genetics and Genomics

    ISSUE

    1-2

    VOLUME

    226

    Author Affiliations

    Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1007/bf00273585

    DOI

    http://dx.doi.org/10.1007/bf00273585

    DIMENSIONS

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    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/2034229


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