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Genetic and physical analyses of Methylobacterium organophilum XX genes encoding methanol oxidation

Journal Article · · J. Bacteriol.; (United States)
OSTI ID:5271834
; ; ;

When allyl alcohol was used as a suicide substrate, spontaneous mutants and UV light- and nitrous acid-generated mutants of Methylobacterium organophilum XX were selected which grew on methylamine but not on methanol. There was no detectable methanol dehydrogenase (MDH) activity in crude extracts of these mutants, yet Western blots revealed that some mutants still produced MDH protein. Complementation of 50 mutants by a cosmid gene bank of M. organophilum XX demonstrated that three major regions of the genome, each of which was separated by a minimum of 40 kilobases, were required for expression of active MDH. By subcloning and Tn5 insertion mutagenesis of subcloned fragments, at least 11 genes clustered within these three regions were subsequently identified. The identity of the MDH structural gene, which was initially determined by hybridization to the structural gene of Methylobacterium sp. strain AM1, was confirmed by Western blot analysis of an MDH-..beta..-galactosidase fusion protein.

Research Organization:
Univ. of Minnesota, Navarre
DOE Contract Number:
AC02-82ER12029
OSTI ID:
5271834
Journal Information:
J. Bacteriol.; (United States), Journal Name: J. Bacteriol.; (United States) Vol. 170:1; ISSN JOBAA
Country of Publication:
United States
Language:
English

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Related Subjects

550400* -- Genetics
560130 -- Radiation Effects on Microorganisms
560300 -- Chemicals Metabolism & Toxicology
59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ALCOHOLS
BACTERIA
CLONING
DNA-CLONING
ELECTROMAGNETIC RADIATION
ENZYMES
GENETIC MAPPING
HYBRIDIZATION
HYDROGEN COMPOUNDS
HYDROXY COMPOUNDS
INORGANIC ACIDS
MAPPING
METHANOL
MICROORGANISMS
MOLECULAR BIOLOGY
MUTAGENESIS
MUTANTS
NITROUS ACID
ORGANIC COMPOUNDS
OXIDOREDUCTASES
RADIATIONS
ULTRAVIOLET RADIATION