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PMID: 6327606 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

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

Journal of bacteriology ·Vol. 158 ·No. 2 ·1984-05-00 ·Pages 488-95

Castilho BA, Olfson P, Casadaban MJ

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.

MeSH Terms
Ampicillin/pharmacology Bacteriophage mu/genetics Chromosome Deletion Chromosomes, Bacterial DNA Transposable Elements DNA, Recombinant Kanamycin/pharmacology Lac Operon Mutation Penicillin Resistance Plasmids Transduction, Genetic
Chemicals
DNA Transposable Elements DNA, Recombinant Kanamycin Ampicillin
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Castilho B A
Olfson P
Casadaban M J
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49 references, click to expand
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.158.2.488-495.1984
Published
1984-05-00
Pages
488-95
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC215454
Subset
IM
Grants
NIAID NIH HHS · AI 00468 · United States
NCI NIH HHS · CA 09273 · United States
NIGMS NIH HHS · GM 29067 · United States
External Links
PubMed source DOI: 10.1128/jb.158.2.488-495.1984 PMC Full Text View journal details
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