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. 1984 May;158(2):488–495. doi: 10.1128/jb.158.2.488-495.1984

Plasmid insertion mutagenesis and lac gene fusion with mini-mu bacteriophage transposons.

B A Castilho, P Olfson, M J Casadaban
PMCID: PMC215454  PMID: 6327606

Abstract

Small bacteriophage Mu transposable elements containing the lac operon structural genes were constructed to facilitate the isolation and use of Mu insertions and lac gene fusions. These mini-Mu elements have selectable genes for either ampicillin or kanamycin resistance and can be used to form both transcriptional and translational lac gene fusions. Some of the mini-Mu-lac elements constructed are deleted for the Mu A and B transposition genes and form stable insertions that cannot undergo transposition unless complemented for these functions. A procedure was developed for selecting mini-Mu insertions specifically into plasmids, including commonly used high-copy-number cloning vectors such as pBR322. Mu insertions in pBR322 were found to be distributed around the plasmid, but insertions in certain regions occurred more frequently than in others.

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Selected References

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