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来自细胞内寄生细菌带绦螺旋体(Caedibacter taeniospiralis)的R体编码质粒pKAP298的序列、转录活性及进化起源

Sequence, transcription activity, and evolutionary origin of the R-body coding plasmid pKAP298 from the intracellular parasitic bacterium Caedibacter taeniospiralis.

作者信息

Jeblick Jörn, Kusch Jürgen

机构信息

Department of Biology/Ecology, University of Kaiserslautern, Erwin-Schroedinger-Strasse 13/14, 67663 Kaiserslautern, Germany.

出版信息

J Mol Evol. 2005 Feb;60(2):164-73. doi: 10.1007/s00239-004-0002-2.

Abstract

We isolated the intracellular parasitic bacterium Caedibacter taeniospiralis from cultures of the freshwater ciliate Paramecium tetraurelia strain 298. Plasmid pKAP298 as well as the total RNA were isolated from the bacteria. pKAP298 was totally sequenced (49.1 kb; NCBI accession number AY422720). From southern blots of pKAP-fragments and Digoxigenin-labeled cDNA of the Caedibacter-RNA, we generated transcription maps of pKAP298. The observed transcription activity indicated functions of the plasmid besides the synthesis of the R-body, a complex protein inclusion associated with toxic effects of Caedibacter cells on host paramecia. We identified 63 potential protein coding regions on pKAP298, and a novel transposon as well as known transposons were characterized. A group II intron was identified. Homologies with putative phage genes were detected on pKAP298 that direct to the evolution of pKAP298 from a bacteriophage. This original phage most probably belonged to the Caudovirales. Hints on a toxin coding region of pKAP298 are given: a protein with homology to the Soj-/ParA-family also has homologies to a membrane associated ATPase, which is involved in eukaryotic ATPase dependent ion carriers and may be associated with toxic effects on paramecia ingesting this protein.

摘要

我们从淡水纤毛虫四膜虫298株的培养物中分离出细胞内寄生细菌带绦螺旋体杆菌。从该细菌中分离出了质粒pKAP298以及总RNA。对pKAP298进行了全序列测定(49.1 kb;NCBI登录号AY422720)。通过pKAP片段的Southern印迹和带绦螺旋体杆菌RNA的地高辛标记cDNA,我们构建了pKAP298的转录图谱。观察到的转录活性表明,除了合成R体(一种与带绦螺旋体杆菌细胞对宿主草履虫的毒性作用相关的复合蛋白包涵体)外,该质粒还具有其他功能。我们在pKAP298上鉴定出63个潜在的蛋白质编码区,并对一个新型转座子以及已知转座子进行了表征。鉴定出一个II类内含子。在pKAP298上检测到与推定噬菌体基因的同源性,这表明pKAP298是由噬菌体进化而来的。这个原始噬菌体很可能属于有尾噬菌体目。文中给出了关于pKAP298毒素编码区的线索:一种与Soj-/ParA-家族具有同源性的蛋白质也与一种膜相关ATP酶具有同源性,该酶参与真核生物依赖ATP酶的离子载体,可能与对摄取这种蛋白质的草履虫的毒性作用有关。

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