1887

Abstract

The genome of strain R has been sequenced completely, but subsequent genetic studies have been limited by the lack of a replicable vector system. In this study, replicable plasmids were constructed for and using the region upstream from the gene. The plasmids of (pGTLori) and (pMIori) replicated in both species, but could not support replication of pGTLori. A 180 bp section of the region of was found to be the minimal region required for plasmid replication in strain S6, the shortest region defined for mycoplasmas. Targeted gene disruption of of S6 was attempted using these plasmids. Constructs made in pPLoriC7 integrated into the genomic region, not into the targeted gene, whereas those made in pMIori disrupted the gene, which has 97 % DNA sequence identity with the gene. During passages, antimicrobial selection pressure did not influence the rate of chromosomal integration. These plasmids will thus be useful for genetic studies, including inactivation or expression of selected genes, in and , and will lead to a better understanding of their molecular biology. They are, to our knowledge, the first replicable plasmids developed for the Pneumoniae phylogenetic group of mycoplasmas.

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2008-09-01
2020-03-30
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