变铅青链霉菌内源线性质粒接合转移必需功能区的鉴定(英文)  

Characterization of The Essential Gene Components for Conjugal Transfer of Streptomyces lividans Linear Plasmid SLP2

作　　者：许铭翾[1] 朱颖旻[1] 沈美娟[1] 姜卫红[1] 赵国屏[1] 覃重军[1,2] 

机构地区：[1]中国科学院上海生命科学院,植物生理生态研究所 [2]Department of Genetics,Stanford University School of Medicine,Stanford,CA 94305-5120,USA

出　　处：《生物化学与生物物理进展》2006年第10期986-993,共8页Progress In Biochemistry and Biophysics

基　　金：国家自然科学基金资助项目(30170019,30270030,30325003,0202ZA14096);国家高技术研究发展计划资助项目(2005AA227020);中国科学院创新项目(KSCX2-SW-329-3)~~

摘　　要：通常细菌间环型质粒在接合转移过程中,单链质粒DNA在质粒内部“oriT”接合转移起始位点发生缺刻.随后,打开的单链质粒DNA通过细胞膜的Ⅳ型分泌系统转移到受体菌中.但是,链霉菌中的接合型线型质粒带有游离3′端,5′端与末端蛋白结合,因而不能以细胞-细胞间方式转移单链缺刻DNA.报道了变铅青链霉菌线型质粒SLP2衍生的环型质粒,与SLP2一样可以高频高效接合转移,并鉴定了接合转移功能区.质粒有效的接合转移功能区包含6个共转录的基因,分别编码一个Tra样的DNA转移酶、胞壁水解酶、2个膜蛋白(可以与ATP结合蛋白相互作用)和一个功能未知的蛋白质.从SalⅠR-/M-向SalⅠR/M宿主转移的质粒频率下降表明,线型和环型的质粒都是以双链的形式转移的.上述研究结果表明SLP2衍生的线型质粒和环型质粒以相似的与细胞膜/胞壁功能相关的机理进行接合转移.Commonly, the interbacterial transfer of circular plasmids is initiated by nicking at an internal sequence, oriT, followed by transferring one strand as single-stranded DNA through a type IV secretion channel on cell membrane. In contrast, Streptomyces conjugative linear plasmids, containing a free 3＇-end but a protein-capped 5＇-end, can potentially undergo cell-to-cell transfer by transfer of non-nicked DNA. It was reported that circular derivatives of the Streptomyces lividans linear plasmid SLP2, as well as the parental linear plasmid itself can transfer efficiently. And the genetic requirements for such transfer was described. Efficient transfer of plasmid requires six co-transcribed SLP2 genes, encoding a Tra-like DNA translocase, cell wall hydrolase, two cell membrane proteins that interact with an ATP binding protein, and a protein of unknown function. Reduced transfer efficiency ofplasmid from Sa/I R-/M- to So/I R/M hosts argues that transfer of both the circular and linear forms of the plasmid involves double-stranded DNA. These results suggest that conjugal transfer occurs by a similar mechanism for SLP2-derived linear and circular plasmids, and cellular membrane/wall functions in the transfer process.

关 键 词：链霉菌 线型质粒 接合转移 

分 类 号：Q93[生物学—微生物学]