Harada Karen Mine, Aso Yuji, Hashimoto Wataru, Mikami Bunzo, Murata Kousaku
Laboratory of Basic and Applied Molecular Biotechnology, Graduate School of Agriculture, Kyoto University, Uji, Kyoto 611-0011, Japan.
Plasmid. 2006 Jul;56(1):11-23. doi: 10.1016/j.plasmid.2005.12.002. Epub 2006 Jan 30.
Sphingomonas sp. A1 (strain A1) is capable of directly incorporating macromolecules (e.g., alginate) through the specialized import system--"super-channel." Here, we report the complete DNA sequence and genetic organization of plasmid pA1 from strain A1. Nucleotide sequence analysis revealed that pA1 comprises 46,557 bp encoding 49 open reading frames (ORFs) with 65% G+C content and abundant GCCG/CGGC motifs. Many predicted pA1 ORFs showed high similarity to pA81 ORFs; pA81 is supposedly a self-transmissible promiscuous incompatibility (Inc) group P-1beta plasmid. Unlike any reported IncP-1 plasmids, pA1 contains no inserted mobile genetic elements. The genetic organization and predicted pA1 ORFs showed greater similarity to the IncP-1beta plasmid backbone than to the IncP-1alpha plasmid backbone. pA1 contains restriction site-associated repeat sequences typical of the IncP-1beta but absent in the IncP-1alpha and delta subgroups. Thus, the overall pA1 structure corresponds to that of the typical IncP-1beta plasmids. Phylogenetic analysis of the replication-associated proteins suggested that pA1 may have diverged later along with the two IncP-1beta plasmids--pA81 and pB4. The 2.4-kb duplicates of stable inheritance genes klcAB and korC in pA1 possibly resulted from insertion and/or recombination events via the repeat sequences flanking these duplicates.
鞘氨醇单胞菌属菌株A1(菌株A1)能够通过特殊的导入系统——“超级通道”直接摄取大分子(如藻酸盐)。在此,我们报告了菌株A1中质粒pA1的完整DNA序列和基因组织。核苷酸序列分析表明,pA1由46,557个碱基对组成,编码49个开放阅读框(ORF),G+C含量为65%,并含有丰富的GCCG/CGGC基序。许多预测的pA1 ORF与pA81 ORF具有高度相似性;pA81据推测是一种自我传递的混杂不相容性(Inc)P-1β组质粒。与任何已报道的IncP-1质粒不同,pA1不包含插入的可移动遗传元件。基因组织和预测的pA1 ORF与IncP-1β质粒骨架的相似性高于与IncP-1α质粒骨架的相似性。pA1含有IncP-1β典型的与限制位点相关的重复序列,但在IncP-1α和δ亚组中不存在。因此,pA1的整体结构与典型的IncP-1β质粒的结构相对应。对复制相关蛋白的系统发育分析表明,pA1可能后来与两个IncP-1β质粒——pA81和pB4一起分化。pA1中稳定遗传基因klcAB和korC的2.4 kb重复序列可能是通过这些重复序列侧翼的重复序列的插入和/或重组事件产生的。