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PMID: 2158981 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Functional analysis of pAL5000, a plasmid from Mycobacterium fortuitum: construction of a "mini" mycobacterium-Escherichia coli shuttle vector.

Journal of bacteriology ·Vol. 172 ·No. 5 ·1990-05-00 ·Pages 2793-7

Ranes MG, Rauzier J, Lagranderie M, Gheorghiu M, Gicquel B

Abstract

Functional domains of pAL5000 were determined by gene disruption and deletion analysis. Of the five plasmid open reading frames (ORFs), ORF1 to ORF5, and a putative origin of replication previously identified (J. Rauzier, J. Moniz-Pereira, and B. Gicquel-Sanzey, Gene 71:315-321), two of the ORFs (ORF3 and ORF4) were deemed dispensable for plasmid replication. A "mini" mycobacterium-Escherichia coli shuttle plasmid applicable for general recombinant DNA studies in mycobacteria was constructed by using the gene for Kanr (Tn903) as a selective marker. Heterologous expression of the gene for Kanr was confirmed by Western blotting (immunoblotting) analysis.

MeSH Terms

Chromosome Deletion DNA Replication DNA Transposable Elements DNA, Bacterial/genetics,isolation & purification Escherichia coli/genetics Gene Expression Genetic Vectors Immunoblotting Mycobacterium/genetics Nucleic Acid Hybridization Plasmids Restriction Mapping Species Specificity

Chemicals

DNA Transposable Elements DNA, Bacterial

Authors & Affiliations

5 authors, click to expand affiliations / ORCID

Ranes M G

Unité de Génie Microbiologique, Institut Pasteur, Paris, France.

Rauzier J

Lagranderie M

Gheorghiu M

Gicquel B

References (10)

10 references, click to expand

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Article Info

Journal

Journal of bacteriology

Abbr.

J Bacteriol

ISSN

0021-9193

DOI

10.1128/jb.172.5.2793-2797.1990

Published

1990-05-00

Pages

2793-7

Language

English

Region

United States

NLM ID

2985120R

PMCID

PMC208931

Subset

External Links

PubMed source DOI: 10.1128/jb.172.5.2793-2797.1990 PMC Full Text View journal details

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Functional analysis of pAL5000, a plasmid from Mycobacterium fortuitum: construction of a "mini" mycobacterium-Escherichia coli shuttle vector.

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