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Journal of Bacteriology, June 2000, p. 3582-3586, Vol. 182, No. 12
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Megaplasmid pRme2011a of Sinorhizobium meliloti Is Not Required for Viability

Ivan J. Oresnik, Shu-Lin Liu, Christopher K. Yost, and Michael F. Hynes*

Department of Biological Sciences, University of Calgary, Calgary, Alberta T2N 1N4, Canada

Received 8 November 1999/Accepted 21 March 2000

We report the curing of the 1,360-kb megaplasmid pRme2011a from Sinorhizobium meliloti strain Rm2011. With a positive selection strategy that utilized Tn5B12-S containing the sacB gene, we were able to cure this replicon by successive rounds of selecting for deletion formation in vivo. Subsequent Southern blot, Eckhardt gel, and pulsed-field gel electrophoresis analyses were consistent with the hypothesis that the resultant strain was indeed missing pRme2011a. The cured derivative grew as well as the wild-type strain in both complex and defined media but was unable to use a number of substrates as a sole source of carbon on defined media.

* Corresponding author. Mailing address: Department of Biological Sciences, University of Calgary, Calgary, Alberta T2N 1N4, Canada. Phone: (403) 220-8473. Fax: (403) 289-9311. E-mail: hynes{at}ucalgary.ca.

Journal of Bacteriology, June 2000, p. 3582-3586, Vol. 182, No. 12
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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