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Methanol Oxidation Genes in the Marine Methanotroph Methylomonas sp. Strain A4

Waechter-Brulla, Daryle and Dispirito, Alan A. and Chistoserdova, Ludmila V. and Lidstrom, Mary E. (1993) Methanol Oxidation Genes in the Marine Methanotroph Methylomonas sp. Strain A4. Journal of Bacteriology, 175 (12). pp. 3767-3775. ISSN 0021-9193. http://resolver.caltech.edu/CaltechAUTHORS:20120308-070527725

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Abstract

Methanol dehydrogenase has been purified from the type I marine methanotroph Methylomonas sp. strain A4 and found to be similar to other methanol dehydrogenase enzymes in subunit composition, molecular mass, and N-terminal sequence of the two subunits. A heterologous gene probe and a homologous oligonucleotide have been used to identify a DNA fragment from Methylomonas sp. strain A4 which contains moxF, the gene encoding the large subunit of methanol dehydrogenase. Protein expression experiments with Escherichia coli, immunoblotting of expression extracts, and partial DNA sequence determination have confirmed the presence of moxF on this DNA fragment. In addition, expression and immunoblot experiments have shown the presence of the genes for the small subunit of methanol dehydrogenase (moxI) and for the methanol dehydrogenase-specific cytochrome c (moxG). The moxG gene product has been shown to be cytochrome c552. The expression experiments have also shown that two other genes are present on this DNA fragment, and our evidence suggests that these are the homologs of moxJ and moxR, whose functions are unknown. Our data suggest that the order of these genes in Methylomonas sp. strain A4 is moxFJGIR, the same as in the facultative methylotrophs. The transcriptional start site for moxF was mapped. The sequence 5' to the transcriptional start does not resemble other promoter sequences, including the putative moxF promoter sequence of facultative methylotrophs. These results suggest that although the order of these genes and the N-terminal amino acid sequence of MoxF and MoxI are conserved between distantly related methylotrophs, the promoters for this gene cluster differ substantially.


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http://jb.asm.org/content/175/12/3767PublisherUNSPECIFIED
Additional Information:© 1993 American Society for Microbiology. Received 18 December 1992. Accepted 12 April 1993. This work was supported by a grant from the Department of Energy (DEFOG0-87ER13753). We thank Stan Tabor for supplying plasmids pGP1-2, pT7-3, pT7-5, and pT7-6.
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Funding AgencyGrant Number
Department of Energy (DOE)DEFOG0-87ER13753
Record Number:CaltechAUTHORS:20120308-070527725
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20120308-070527725
Official Citation:Methanol oxidation genes in the marine methanotroph Methylomonas sp. strain A4. D Waechter-Brulla, A A DiSpirito, L V Chistoserdova, and M E Lidstrom J. Bacteriol. June 1993 175:3767-3775
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:29637
Collection:CaltechAUTHORS
Deposited By: Ruth Sustaita
Deposited On:12 Mar 2012 21:08
Last Modified:12 Mar 2012 21:08

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