Construction of broad-host-range cosmid cloning vectors: identification of genes necessary for growth of Methylobacterium organophilum on methanol
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.
- Research Organization:
- Gray Freshwater Biological Institute, Navarre, MI
- DOE Contract Number:
- AC02-82ER12029
- OSTI ID:
- 5614504
- Journal Information:
- J. Bacteriol.; (United States), Journal Name: J. Bacteriol.; (United States) Vol. 161:3; ISSN JOBAA
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
560302 -- Chemicals Metabolism & Toxicology-- Microorganisms-- (-1987)
59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ALCOHOLS
ANTI-INFECTIVE AGENTS
ANTIBIOTICS
BACTERIA
CLONING
DNA
DRUGS
ENZYMES
GENES
GENOME MUTATIONS
GROWTH
HYDROXY COMPOUNDS
METABOLISM
METHANOGENIC BACTERIA
METHANOL
MICROORGANISMS
MUTANTS
MUTATIONS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
OXIDOREDUCTASES
RECOMBINANT DNA