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Applied and Environmental Microbiology, February 1999, p. 648-651, Vol. 65, No. 2
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Detection of Methanotrophs in Groundwater by PCRdagger

Y. S. Cheng, J. L. Halsey, K. A. Fode, C. C. Remsen, and M. L. P. Collins*

Department of Biological Sciences and Great Lakes Water Institute, University of Wisconsin---Milwaukee, Milwaukee, Wisconsin 53201

Received 1 October 1998/Accepted 16 November 1998

Methanotrophic bacteria have significant potential for bioremediation, which would require methods for monitoring the presence and activity of these organisms in environmental samples. In this study, PCR was used to detect methanotrophic bacteria. Primers were designed on the basis of a partial sequence of pmoA, which encodes one of the proteins of the particulate methane monooxygenase. Specific amplification of a portion of pmoA was obtained with template DNA isolated from lab strains of methanotrophs. A pmoA product was also obtained by using DNA from groundwater. The identity of the PCR product was confirmed by sequencing or by amplification with a nested primer. Reverse transcriptase PCR detected pmoA mRNA.

* Corresponding author. Mailing address: Department of Biological Sciences, University of Wisconsin, Milwaukee, P.O. Box 413, Milwaukee, WI 53201. Phone: (414) 229-5298. Fax: (414) 229-3926. E-mail: mlpcolli{at}uwm.edu.

dagger This is publication no. 416 from the Center for Great Lakes Studies.

Applied and Environmental Microbiology, February 1999, p. 648-651, Vol. 65, No. 2
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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