Plasmid replication region
Springer Online Journal Archives 1860-2000
Summary Derivatives of the conjugative streptococcal plasmid pIP501 replicate stably in Bacillus subtilis. The region essential for replication of plP501 has been narrowed down to a 2.2 kb DNA segment, the sequence of which has been determined. This region comprises two genes, copR and repR, proposed to be involved in copy control and replication. By in vitro and in vivo transcriptional analysis we characterized three active promoters, pII pII and pIII within this region. A putative fourth promoter (PTV) was neither active in vitro nor in vivo. We showed that copR is transcribed from promoter pI while the repR, gene is transcribed from promoter pII located just downstream of copR The pII transcript encompasses a 329 nucleotide (nt) long leader sequence. A counter transcript that was complementary to a major part of this leader was found to originate from a third promoter pIII The secondary structure of the counter transcript revealed several stem-loop regions. A regulatory function for this antisense RNA in the control of repR, expression is proposed. Comparative analysis of the replication regions of pAMβ 1 and pSM19035 suggested a similar organization of transcriptional units, suggesting that an antisense RNA is produced by these plasmids also.
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