In vitro and in vivo analysis of transcription within the replication region of plasmid pIP501
- 1 July 1992
- journal article
- research article
- Published by Springer Nature in Molecular Genetics and Genomics
- Vol. 234 (1) , 105-112
- https://doi.org/10.1007/bf00272351
Abstract
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.Keywords
This publication has 45 references indexed in Scilit:
- Structurally stable Bacillus subtilis cloning vectorsGene, 1990
- Segregational instability of pUB110-derived recombinant plasmids in Bacillus subtilisPlasmid, 1985
- Double or triple sets of replication functions as inverted and direct repeats onin vitro reconstructed streptococcal MLS resistance plasmidsJournal of Basic Microbiology, 1983
- Location of antibiotic resistance determinants, copy control, and replication functions on the double-selective streptococcal cloning vector pGB301Molecular Genetics and Genomics, 1981
- Plasmid pGB301, a new multiple resistance streptococcal cloning vehicle and its use in cloning of a gentamicin/kanamycin resistance determinantMolecular Genetics and Genomics, 1981
- Physical mapping of plasmid pDB101: A potential vector plasmid for molecular cloning in streptococciPlasmid, 1980
- Electron microscopic mapping of deletions on a streptococcal plasmid carrying extraordinarily long inverted repeatsPlasmid, 1980
- Post-transformational rearrangement of an in vitro reconstructed group-A streptococcal erythromycin resistance plasmidPlasmid, 1979
- RNA Polymerase binding sites and transcription map of the DNA of Bacillus subtilis phage φ29Journal of Molecular Biology, 1979
- R Plasmids in Streptococcus agalactiae (Group B)Antimicrobial Agents and Chemotherapy, 1976