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  • 1
    ISSN: 1432-072X
    Keywords: Transposon5 ; Xylose operon ; Transcriptional fusions ; ColE1 replication ; luxAB genes ; Escherichia coli ; Vibrio harveyi
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A ColE1-based plasmid for transcriptional gene fusions was constructed that contains both the promoterless luxAB genes of Vibrio harveyi and a tet marker within the inverted repeats of a left end-truncated Tn5 element. Introduction of this plasmid into an Escherichia coli strain containing a plasmid (pTF421) that overproduces ColE1 RNA1 (and thus inhibits replication of the ColE1 plasmid) allowed selection for cells that had a single copy of the luxAB operon transposed into the chromosome beginning 5 days post-transformation. The long latent period necessary for Tn5 transposition is analogous to that found in other systems, where transposition frequencies and mutation rates increase in a time-dependent manner when selected for upon prolonged incubation on Petri dishes under bacteriostatic conditions.
    Type of Medium: Electronic Resource
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