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广宿主范围IncQ样质粒pTC-F14的转移区域及其与相关质粒pTF-FC2相互作用能力的分析

Analysis of the mobilization region of the broad-host-range IncQ-like plasmid pTC-F14 and its ability to interact with a related plasmid, pTF-FC2.

作者信息

van Zyl Leonardo J, Deane Shelly M, Rawlings Douglas E

机构信息

Department of Microbiology, University of Stellenbosch, Matieland 7600, South Africa.

出版信息

J Bacteriol. 2003 Oct;185(20):6104-11. doi: 10.1128/JB.185.20.6104-6111.2003.

Abstract

Plasmid pTC-F14 is a 14.2-kb plasmid isolated from Acidithiobacillus caldus that has a replicon that is closely related to the promiscuous, broad-host-range IncQ family of plasmids. The region containing the mobilization genes was sequenced and encoded five Mob proteins that were related to those of the DNA processing (Dtr or Tra1) region of IncP plasmids rather than to the three-Mob-protein system of the IncQ group 1 plasmids (e.g., plasmid RSF1010 or R1162). Plasmid pTC-F14 is the second example of an IncQ family plasmid that has five mob genes, the other being pTF-FC2. The minimal region that was essential for mobilization included the mobA, mobB, and mobC genes, as well as the oriT gene. The mobD and mobE genes were nonessential, but together, they enhanced the mobilization frequency by approximately 300-fold. Mobilization of pTC-F14 between Escherichia coli strains by a chromosomally integrated RP4 plasmid was more than 3,500-fold less efficient than the mobilization of pTF-FC2. When both plasmids were coresident in the same E. coli host, pTC-F14 was mobilized at almost the same frequency as pTF-FC2. This enhanced pTC-F14 mobilization frequency was due to the presence of a combination of the pTF-FC2 mobD and mobE gene products, the functions of which are still unknown. Mob protein interaction at the oriT regions was unidirectionally plasmid specific in that a plasmid with the oriT region of pTC-F14 could be mobilized by pTF-FC2 but not vice versa. No evidence for any negative effect on the transfer of one plasmid by the related, potentially competitive plasmid was obtained.

摘要

质粒pTC-F14是从嗜热嗜酸硫杆菌中分离出的一个14.2 kb的质粒,其复制子与混杂的、广宿主范围的IncQ质粒家族密切相关。对包含转移基因的区域进行了测序,该区域编码了5种Mob蛋白,这些蛋白与IncP质粒的DNA加工(Dtr或Tra1)区域的蛋白相关,而不是与IncQ 1组质粒(如质粒RSF1010或R1162)的三Mob蛋白系统相关。质粒pTC-F14是具有5个mob基因的IncQ家族质粒的第二个例子,另一个是pTF-FC2。转移所必需的最小区域包括mobA、mobB和mobC基因以及oriT基因。mobD和mobE基因不是必需的,但它们共同作用可使转移频率提高约300倍。通过染色体整合的RP4质粒在大肠杆菌菌株之间转移pTC-F14的效率比转移pTF-FC2低3500倍以上。当这两种质粒共存于同一大肠杆菌宿主中时,pTC-F14的转移频率几乎与pTF-FC2相同。pTC-F14转移频率的提高是由于存在pTF-FC2的mobD和mobE基因产物的组合,其功能仍不清楚。oriT区域的Mob蛋白相互作用在质粒特异性上是单向的,即具有pTC-F14 oriT区域的质粒可以被pTF-FC2转移,但反之则不行。没有获得相关的、可能具有竞争性的质粒对一种质粒转移产生任何负面影响的证据。

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