Nucleotide sequence determination and genetic analysis of the Bacteroides plasmid, pBI143.

The nucleotide sequence and genetic organization of the Bacteroides plasmid pBI143 were determined. The plasmid was 2747 base pairs (bp) and had a G+C content of 41% (GenBank Accession No. U30316). There were two open reading frames greater than 50 codons and these were designated mobA and repA. A 56-bp inverted repeat divided pBI143 into modules with repA and mobA in separate regions. There was a marked difference in the G+C content and codon usage for the two regions; repA had 33% G+C and mobA was 44% G+C. MobA had homology to other Bacteroides mobilization proteins and RepA shared homology to a replication protein from Zymomonas mobilis plasmid pZM2. These two putative replication proteins formed a subgroup of the rolling-circle replication.proteins belonging to the pSN2 family of gram-positive plasmids. Consistent with this finding, single-stranded pBI143 DNA was detected in plasmid containing Bacteroides fragilis cultures. Availability of the pBI143 sequence allowed the elucidation of the complete nucleotide sequence for pFD288 an 8.9-kb Bacteroides shuttle vector (GenBank Accession No. U30830).