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The particulate methane monooxygenase gene clusters, pmoCAB, from two representative type II methanotrophs of the alpha-Proteobacteria, Methylosinus trichosporium OB3b and Methylocystis sp. strain M, have been cloned and sequenced. Primer extension experiments revealed that the pmo cluster is probably transcribed from a single transcriptional start site located 300 bp upstream of the start of the first gene, pmoC, for Methylocystis sp. strain M. Immediately upstream of the putative start site, consensus sequences for sigma(70) promoters were identified, suggesting that these pmo genes are recognized by sigma(70) and negatively regulated under low-copper conditions. The pmo genes were cloned in several overlapping fragments, since parts of these genes appeared to be toxic to the Escherichia coli host. Methanotrophs contain two virtually identical copies of pmo genes, and it was necessary to use Southern blotting and probing with pmo gene fragments in order to differentiate between the two pmoCAB clusters in both methanotrophs. The complete DNA sequence of one copy of pmo genes from each organism is reported here. The gene sequences are 84% similar to each other and 75% similar to that of a type I methanotroph of the gamma-Proteobacteria, Methylococcus capsulatus Bath. The derived proteins PmoC and PmoA are predicted to be highly hydrophobic and consist mainly of transmembrane-spanning regions, whereas PmoB has only two putative transmembrane-spanning helices. Hybridization experiments showed that there are two copies of pmoC in both M. trichosporium OB3b and Methylocystis sp. strain M, and not three copies as found in M. capsulatus Bath.
B Gilbert, and
I R McDonald, and
R Finch, and
G P Stafford, and
A K Nielsen, and
J C Murrell
May 2004,
Applied and environmental microbiology,
B Gilbert, and
I R McDonald, and
R Finch, and
G P Stafford, and
A K Nielsen, and
J C Murrell
June 1995,
Journal of bacteriology,
B Gilbert, and
I R McDonald, and
R Finch, and
G P Stafford, and
A K Nielsen, and
J C Murrell
January 1999,
Archives of microbiology,
B Gilbert, and
I R McDonald, and
R Finch, and
G P Stafford, and
A K Nielsen, and
J C Murrell
September 2022,
Nature communications,
B Gilbert, and
I R McDonald, and
R Finch, and
G P Stafford, and
A K Nielsen, and
J C Murrell
January 2004,
Marine biotechnology (New York, N.Y.),
B Gilbert, and
I R McDonald, and
R Finch, and
G P Stafford, and
A K Nielsen, and
J C Murrell
February 2011,
Environmental microbiology reports,
B Gilbert, and
I R McDonald, and
R Finch, and
G P Stafford, and
A K Nielsen, and
J C Murrell
March 2009,
Wei sheng wu xue bao = Acta microbiologica Sinica,
B Gilbert, and
I R McDonald, and
R Finch, and
G P Stafford, and
A K Nielsen, and
J C Murrell
June 2009,
Applied biochemistry and biotechnology,
B Gilbert, and
I R McDonald, and
R Finch, and
G P Stafford, and
A K Nielsen, and
J C Murrell
January 2011,
Methods in enzymology,
B Gilbert, and
I R McDonald, and
R Finch, and
G P Stafford, and
A K Nielsen, and
J C Murrell
November 1997,
FEMS microbiology letters,
