Megaplasmid pRme2011a of Sinorhizobium meliloti is not required for viability

J Bacteriol. 2000 Jun;182(12):3582-6. doi: 10.1128/JB.182.12.3582-3586.2000.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blotting, Southern
  • Electrophoresis, Gel, Pulsed-Field
  • Gene Deletion
  • Genetic Markers
  • Phenotype
  • Plasmids / genetics*
  • Sinorhizobium meliloti / genetics*
  • Sinorhizobium meliloti / growth & development*

Substances

  • Genetic Markers