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Identification of putative methanol dehydrogenase (moxF) structural genes in methylotrophs and cloning of moxF genes from Methylococcus capsulatus bath and Methylomonas albus BG8

Stephens, Richard L. and Haygood, Margo G. and Lidstrom, Mary E. (1988) Identification of putative methanol dehydrogenase (moxF) structural genes in methylotrophs and cloning of moxF genes from Methylococcus capsulatus bath and Methylomonas albus BG8. Journal of Bacteriology, 170 (5). pp. 2063-2069. ISSN 0021-9193. http://resolver.caltech.edu/CaltechAUTHORS:STEjbact88

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Abstract

An open-reading-frame fragment of a Methylobacterium sp. strain AM1 gene (moxF) encoding a portion of the methanol dehydrogenase structural protein has been used as a hybridization probe to detect similar sequences in a variety of methylotrophic bacteria. This hybridization was used to isolate clones containing putative moxF genes from two obligate methanotrophic bacteria, Methylococcus capsulatus Bath and Methylomonas albus BG8. The identity of these genes was confirmed in two ways. A T7 expression vector was used to produce methanol dehydrogenase protein in Escherichia coli from the cloned genes, and in each case the protein was identified by immunoblotting with antiserum against the Methylomonas albus methanol dehydrogenase. In addition, a moxF mutant of Methylobacterium strain AM1 was complemented to a methanol-positive phenotype that partially restored methanol dehydrogenase activity, using broad-host-range plasmids containing the moxF genes from each methanotroph. The partial complementation of a moxF mutant in a facultative serine pathway methanol utilizer by moxF genes from type I and type X obligate methane utilizers suggests broad functional conservation of the methanol oxidation system among gram-negative methylotrophs.


Item Type:Article
Additional Information:Copyright © 1988 American Society for Microbiology. Received 20 October 1987. Accepted 10 February 1988. This research was supported by Public Health Service grant GM36296 from the National Institutes of Health. We thank David Nunn for providing the Methylomonas albus BG8 clone bank, Lorie Buchholz for providing the Methylomonas albus BG8 MeDH antiserum, and Dawn Anderson for developing the Methylobacterium strain AM1 expression protocol.
Record Number:CaltechAUTHORS:STEjbact88
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:STEjbact88
Alternative URL:http://jb.asm.org/cgi/content/abstract/170/5/2063
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:2947
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:08 May 2006
Last Modified:26 Dec 2012 08:51

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