Plasmid analysis and cloning of the dichloromethane-utilization genes of Methylobacterium sp. DM4.
J Gen Microbiol
; 134(4): 943-52, 1988 Apr.
Article
en En
| MEDLINE
| ID: mdl-3141582
ABSTRACT
The dichloromethane (DCM)-utilizing facultative methylotroph Methylobacterium sp. DM4 was shown to contain three plasmids with approximate size of 120 kb, 40 kb and 8 kb. Curing experiments suggested that the DCM-utilization character was correlated with the possession of an intact 120 kb plasmid. The DCM-utilization genes were cloned on the broad-host-range vector pVK100. Plasmid pME1510, a recombinant plasmid carrying a 21 kb HindIII fragment complemented DCM-utilization-negative derivatives of Methylobacterium sp. DM4 and conferred the DCM-utilization-positive phenotype to a number of Gram-negative methylotrophic bacteria. In Southern hybridization experiments with pMe1510 as a probe, chromosomal DNA from Methylobacterium sp. DM4 gave definite signals while purified plasmid DNA did not. Plasmid pME1510 did not hybridize with total DNA from a cured DCM-non-utilizing derivative of Methylobacterium sp. DM4. It is concluded that the DCM-utilization genes are located on the chromosome or on a megaplasmid. Curing procedures thus led to the formation of a chromosomal or megaplasmid deletion larger than 21 kb and covering the DCM-utilization genes or to the loss of an undetected megaplasmid.
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Banco de datos:
MEDLINE
Asunto principal:
Plásmidos
/
Clonación Molecular
/
Methylococcaceae
/
Genes Bacterianos
Idioma:
En
Año:
1988
Tipo del documento:
Article