This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Gilbert, B. Articles by Murrell, J. C. Search for Related Content PubMed PubMed Citation Articles by Gilbert, B. Articles by Murrell, J. C. Agricola Articles by Gilbert, B. Articles by Murrell, J. C.

Applied and Environmental Microbiology, March 2000, p. 966-975, Vol. 66, No. 3
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Molecular Analysis of the pmo (Particulate Methane Monooxygenase) Operons from Two Type II Methanotrophs

Bettina Gilbert, Ian R. McDonald, Ruth Finch, Graham P. Stafford, Allan K. Nielsen, and J. Colin Murrell^*

Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, United Kingdom

Received 11 October 1999/Accepted 14 December 1999

The particulate methane monooxygenase gene clusters, pmoCAB, from two representative type II methanotrophs of the -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 ⁷⁰ promoters were identified, suggesting that these pmo genes are recognized by 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 -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.

---

^* Corresponding author. Mailing address: Department of Biological Sciences, University of Warwick, Coventry, CV4 7AL, United Kingdom. Phone: 44 (0) 2476 523553. Fax: 44 (0) 2476 523568. E-mail: cm{at}dna.bio.warwick.ac.uk.

Present address: Horticulture Research International, Wellesbourne, Warwick CV35 9EF, United Kingdom.

Present address: Department of Microbiology, Technical University of Denmark, DK-2800 Lyngby, Denmark.

---

Applied and Environmental Microbiology, March 2000, p. 966-975, Vol. 66, No. 3
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

This article has been cited by other articles:

• McDonald, I. R., Bodrossy, L., Chen, Y., Murrell, J. C. (2008). Molecular Ecology Techniques for the Study of Aerobic Methanotrophs. Appl. Environ. Microbiol. 74: 1305-1315 [Full Text]  
• Knapp, C. W., Fowle, D. A., Kulczycki, E., Roberts, J. A., Graham, D. W. (2007). From the Cover: Methane monooxygenase gene expression mediated by methanobactin in the presence of mineral copper sources. Proc. Natl. Acad. Sci. USA 104: 12040-12045 [Abstract] [Full Text]  
• Ali, H., Scanlan, J., Dumont, M. G., Murrell, J. C. (2006). Duplication of the mmoX gene in Methylosinus sporium: cloning, sequencing and mutational analysis.. Microbiology 152: 2931-2942 [Abstract] [Full Text]  
• Ricke, P., Kube, M., Nakagawa, S., Erkel, C., Reinhardt, R., Liesack, W. (2005). First Genome Data from Uncultured Upland Soil Cluster Alpha Methanotrophs Provide Further Evidence for a Close Phylogenetic Relationship to Methylocapsa acidiphila B2 and for High-Affinity Methanotrophy Involving Particulate Methane Monooxygenase. Appl. Environ. Microbiol. 71: 7472-7482 [Abstract] [Full Text]  
• Borodina, E., McDonald, I. R., Murrell, J. C. (2004). Chloromethane-Dependent Expression of the cmu Gene Cluster of Hyphomicrobium chloromethanicum. Appl. Environ. Microbiol. 70: 4177-4186 [Abstract] [Full Text]  
• Ricke, P., Erkel, C., Kube, M., Reinhardt, R., Liesack, W. (2004). Comparative Analysis of the Conventional and Novel pmo (Particulate Methane Monooxygenase) Operons from Methylocystis Strain SC2. Appl. Environ. Microbiol. 70: 3055-3063 [Abstract] [Full Text]  
• Tchawa Yimga, M., Dunfield, P. F., Ricke, P., Heyer, J., Liesack, W. (2003). Wide Distribution of a Novel pmoA-Like Gene Copy among Type II Methanotrophs, and Its Expression in Methylocystis Strain SC2. Appl. Environ. Microbiol. 69: 5593-5602 [Abstract] [Full Text]  
• Stafford, G. P., Scanlan, J., McDonald, I. R., Murrell, J. C. (2003). rpoN, mmoR and mmoG, genes involved in regulating the expression of soluble methane monooxygenase in Methylosinus trichosporium OB3b. Microbiology 149: 1771-1784 [Abstract] [Full Text]  
• Csaki, R., Bodrossy, L., Klem, J., Murrell, J. C., Kovacs, K. L. (2003). Genes involved in the copper-dependent regulation of soluble methane monooxygenase of Methylococcus capsulatus (Bath): cloning, sequencing and mutational analysis. Microbiology 149: 1785-1795 [Abstract] [Full Text]  
• Baxter, N. J., Scanlan, J., De Marco, P., Wood, A. P., Murrell, J. C. (2002). Duplicate Copies of Genes Encoding Methanesulfonate Monooxygenase in Marinosulfonomonas methylotropha Strain TR3 and Detection of Methanesulfonate Utilizers in the Environment. Appl. Environ. Microbiol. 68: 289-296 [Abstract] [Full Text]  
• Bourne, D. G., McDonald, I. R., Murrell, J. C. (2001). Comparison of pmoA PCR Primer Sets as Tools for Investigating Methanotroph Diversity in Three Danish Soils. Appl. Environ. Microbiol. 67: 3802-3809 [Abstract] [Full Text]  
• Stolyar, S., Franke, M., Lidstrom, M. E. (2001). Expression of Individual Copies of Methylococcus capsulatus Bath Particulate Methane Monooxygenase Genes. J. Bacteriol. 183: 1810-1812 [Abstract] [Full Text]  
• Auman, A. J., Stolyar, S., Costello, A. M., Lidstrom, M. E. (2000). Molecular Characterization of Methanotrophic Isolates from Freshwater Lake Sediment. Appl. Environ. Microbiol. 66: 5259-5266 [Abstract] [Full Text]