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PMID: 2982796 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S.

Construction of broad-host-range cosmid cloning vectors: identification of genes necessary for growth of Methylobacterium organophilum on methanol.

Journal of bacteriology ·Vol. 161 ·No. 3 ·1985-03-00 ·Pages 955-62

Allen LN, Hanson RS

Abstract

Four new cloning vectors have been constructed from the broad-host-range cloning vector pRK290. These vectors, pLA2901, pLA2905, pLA2910, and pLA2917, confer resistance to kanamycin and tetracycline. The latter two are cosmid derivatives of pLA2901. The new vectors can be mobilized into, and are stably maintained in, a variety of gram-negative bacteria. A Sau3A genomic bank of Methylobacterium organophilum strain xx DNA has been constructed in pLA2917, and complementation analysis, with a variety of mutants unable to grow on methanol, revealed at least five separate regions necessary for growth on methanol. Complementation analysis and Tn5 mutagenesis data suggest that at least three genes are responsible for expression of active methanol dehydrogenase.

MeSH Terms
Carbon/metabolism Cloning, Molecular DNA Restriction Enzymes/biosynthesis Genetic Complementation Test Genetic Vectors Methanol/metabolism Methylococcaceae/genetics,growth & development
Chemicals
Carbon DNA Restriction Enzymes Methanol
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Allen L N
Hanson R S
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21 references, click to expand
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.161.3.955-962.1985
Published
1985-03-00
Pages
955-62
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC214991
Subset
IM
External Links
PubMed source DOI: 10.1128/jb.161.3.955-962.1985 PMC Full Text View journal details
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