Detection of soluble methane monooxygenase producing Methylosinus trichosporium OB3b by polymerase chain reaction.
Can J Microbiol
; 40(11): 969-73, 1994 Nov.
Article
em En
| MEDLINE
| ID: mdl-7804907
ABSTRACT
The soluble methane monooxygenase (sMMO) enzyme complex of methanotrophs cometabolizes haloaliphatic compounds such as trichloroethylene. Two 18-mer oligonucleotides as primary primers and a nested primer of the same length were selected to amplify specific DNA sequences of the sMMO gene cluster using polymerase chain reaction (PCR). Two DNA fragments of sizes 270 and 400 base pairs were obtained when purified DNA from the methanotroph Methylosinus trichosporium OB3b was used as template. The primers were specific for sMMO sequences of M. trichosporium, since none of the 13 bacterial isolates screened yielded the expected length of PCR-amplified DNA fragments. The detection limit of the PCR method was 5 x 10(2) cells of M. trichosporium. The sMMO sequences were successfully amplified in groundwater (containing native microbial population) when seeded with M. trichosporium, FP1 sense (5'-ATGTCCAGCGCTCATAAC-3'), RP1 antisense (5'-TCAGATGTCGGTCAGGGC-3'), FP2 sense nested (5'GCCATCATCGGTCAGGGC-3'), and FP2 sense nested (5'-GCCATCATCGAGGACATC-3').
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Oxigenases
/
Microbiologia da Água
/
DNA Bacteriano
/
Reação em Cadeia da Polimerase
/
Methylococcaceae
Tipo de estudo:
Diagnostic_studies
Idioma:
En
Revista:
Can J Microbiol
Ano de publicação:
1994
Tipo de documento:
Article