Cloning and analysis of a large plasmid pBMB165 from Bacillus thuringiensis revealed a novel plasmid organization

PLoS One. 2013 Dec 2;8(12):e81746. doi: 10.1371/journal.pone.0081746. eCollection 2013.

Abstract

In this study, we report a rapid cloning strategy for large native plasmids via a contig linkage map by BAC libraries. Using this method, we cloned a large plasmid pBMB165 from Bacillus thuringiensis serovar tenebrionis strain YBT-1765. Complete sequencing showed that pBMB165 is 77,627 bp long with a GC-content of 35.36%, and contains 103 open reading frames (ORFs). Sequence analysis and comparison reveals that pBMB165 represents a novel plasmid organization: it mainly consists of a pXO2-like replicon and mobile genetic elements (an inducible prophage BMBTP3 and a set of transposable elements). This is the first description of this plasmid organization pattern, which may result from recombination events among the plasmid replicon, prophage and transposable elements. This plasmid organization reveals that the prophage BMBTP3 may use the plasmid replicon to maintain its genetic stability. Our results provide a new approach to understanding co-evolution between bacterial plasmids and bacteriophage.

MeSH terms

  • Bacillus thuringiensis / genetics*
  • Bacillus thuringiensis / virology
  • Chromosome Mapping
  • Chromosomes, Artificial, Bacterial / genetics
  • Cloning, Molecular
  • Evolution, Molecular
  • Plasmids / genetics*
  • Prophages / genetics
  • Prophages / physiology
  • Sequence Analysis, DNA

Grants and funding

This work was supported by National High Technology Research and Development Program (863 Program) of China (Grant 2011AA10A203), National Basic Research Program (973 Program) of China (Grant 2009CB118902), National Natural Science Foundation of China (Grants 31000020 and 31170047), and China 948 Program of Ministry of Agriculture (2011-G25).The website is: http://program.most.gov.cn/. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.