State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University, Wuhan, Hubei, People's Republic of China.
PLoS One. 2013 Dec 2;8(12):e81746. doi: 10.1371/journal.pone.0081746. eCollection 2013.
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.
在这项研究中，我们通过 BAC 文库报道了一种通过连续图谱快速克隆天然大质粒的策略。使用该方法，我们从苏云金芽孢杆菌 serovar tenebrionis 菌株 YBT-1765 中克隆了一个大质粒 pBMB165。完整的测序表明，pBMB165 长 77627bp，GC 含量为 35.36%，包含 103 个开放阅读框 (ORFs)。序列分析和比较表明，pBMB165 代表了一种新的质粒组织形式：它主要由 pXO2 样复制子和移动遗传元件（一个诱导性原噬菌体 BMBTP3 和一组转座元件）组成。这是对这种质粒组织形式的首次描述，可能是质粒复制子、原噬菌体和转座元件之间重组事件的结果。这种质粒组织形式表明，原噬菌体 BMBTP3 可能利用质粒复制子来维持其遗传稳定性。我们的研究结果为理解细菌质粒和噬菌体之间的共同进化提供了新的途径。
