Tani Akio, Tanaka Akiyuki, Minami Toshiyuki, Kimbara Kazuhide, Kawai Fusako
Institute of Plant Science and Resources, Okayama University, Okayama, Japan.
Biosci Biotechnol Biochem. 2011;75(2):295-8. doi: 10.1271/bbb.100650. Epub 2011 Feb 7.
A cryptic plasmid, pSM103mini, was found in polyethylene-glycol degrading bacterium Sphingopyxis macrogoltabida, strain 103. The plasmid was 4,328-bp long and its GC content was 57.5%. It contained four open reading frames, but only two of them showed significant similarity to reported proteins. ORF3 and ORF4 were assumed to encode resolvase and replication protein (RepA) respectively. Downstream of ORF4 we found complex repeat sequences. These together with ORF3 and 4 were necessary and sufficient for plasmid maintenance in strain 103, as analyzed by constructing deletion plasmids. The pHSG398-fused plasmid (pHSG-SM103mini) functioned as a shuttle vector between strain 103 and Escherichia coli. The plasmid constructed was maintained in strain 103 and its close relative, S. macrogoltabida strain 203, but not efficiently in PEG-degrading S. terrae.
在聚乙二醇降解菌大谷鞘氨醇单胞菌(Sphingopyxis macrogoltabida)103菌株中发现了一种隐蔽质粒pSM103mini。该质粒长度为4328 bp,GC含量为57.5%。它包含四个开放阅读框,但其中只有两个与已报道的蛋白质有显著相似性。假定ORF3和ORF4分别编码解离酶和复制蛋白(RepA)。在ORF4下游我们发现了复杂的重复序列。通过构建缺失质粒分析发现,这些序列与ORF3和ORF4一起对于103菌株中的质粒维持是必要且充分的。pHSG398融合质粒(pHSG-SM103mini)在103菌株和大肠杆菌之间起穿梭载体的作用。构建的质粒在103菌株及其近缘菌株大谷鞘氨醇单胞菌203菌株中得以维持,但在聚乙二醇降解菌土壤鞘氨醇单胞菌(S. terrae)中维持效率不高。