首页> 外文期刊>Plasmid: An International Journal Devoted to Extrachromosomal Gene Systems >Identification of the origin of replications and partial characterization of plasmid pRK100.
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Identification of the origin of replications and partial characterization of plasmid pRK100.

机译:复制起点的鉴定和质粒pRK100的部分表征。

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摘要

In search for the evolutionary origin of the conjugative F-like plasmid pRK100, the plasmid's functional replication regions were identified. Additionally targeted genetic analysis was used to investigate origins of other regions of the plasmid. Construction of minireplicons via ligation of Tn1725 with plasmid fragments and targeted cloning of putative replication regions, followed by sequence analysis indicated two functional replication regions, a F plasmid related RepFIB and a R1 plasmid related RepFIIA replication region. Partial nucleotide sequencing of regions of the plasmid revealed genes that encode a putative enterochelin iron uptake system previously associated with an Escherichia coli pathogenicity island, PAI III(536), and the pColV-like aerobactin genes. In addition, a homologue of the R100 plasmid related rmoA gene was found that exhibits strong similarity to hha/ymoA encoding the Hha/YmoA class of modulators of gene expression. PCR and hybridization experiments further demonstrated that pRK100 harbors multiple IS2 and IS3 insertion sequences that may have facilitated in the acquisition of elements from other DNA molecules. These data together with the previous identification of a F-like tra region and a pColIa-like colicin Ia, indicate that pRK100 has a highly mosaic structure with elements derived from many different known large natural plasmids.
机译:为了寻找偶联的F样质粒pRK100的进化起源,鉴定了质粒的功能复制区。另外,有针对性的遗传分析用于研究质粒其他区域的起源。通过将Tn1725与质粒片段连接并靶向克隆假定的复制区,然后进行序列分析,发现了两个微型复制区,一个F质粒相关的RepFIB和一个R1质粒相关的RepFIIA复制区。质粒区域的部分核苷酸测序揭示了编码假定的肠螯合铁摄取系统的基因,该系统先前与大肠杆菌的致病性岛,PAI III(536)和pColV样的航空细菌素基因有关。另外,发现与R100质粒相关的rmoA基因的同源物与编码基因表达的Hha / YmoA类调节剂的hha / ymoA表现出高度相似性。 PCR和杂交实验进一步证明,pRK100具有多个IS2和IS3插入序列,这些序列可能有助于从其他DNA分子中获取元素。这些数据以及先前对F样tra区域和pColIa样大肠菌素Ia的鉴定表明,pRK100具有高度镶嵌的结构,其元素衍生自许多不同的已知大型天然质粒。

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