鳗弧菌RTX基因簇的克隆及相关基因功能的初步研究
首发时间:2007-10-17
摘要:以鳗弧菌(Vibrio anguillarum)野生株MVM425基因组DNA为模板,根据已报道的弧菌属I型分泌系统相关基因的保守序列设计引物,通过PCR及genome walking等方法,克隆得到长约9 kb的鳗弧菌I型分泌系统RTX基因簇,包括三个完整分泌元件基因rtxB,rtxC,rtxD,及部分被分泌蛋白编码基因rtxA。为了考察上述系统的分泌功能,利用插入失活技术分别构建得到rtxB,rtxC的突变株VABM和VACM,选取鳗弧菌的两类重要分泌型物质(胞外多糖和胞外蛋白酶)为研究对象,检测结果表明:与野生株相比,突变株的胞外多糖活性无明显变化,胞外总蛋白酶活性分别降低71%(VABM)和58%(VACM)。因此,鳗弧菌I型分泌系统可能参与了部分蛋白酶的分泌,而没有参与多糖类物质的胞外运输过程。
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Cloning and characterization of the RTX cluster in Vibrio anguillarum
Abstract:A 9 kb fragment was cloned from wild type Vibrio anguillarum MVM425 genome DNA by PCR. Sequencing data indicated that this fragment encoded a RTX cluster which was regarded as a typical member of type I secretion system, including three ORFs rtxB, rtxC, rtxD and partial of a putative toxin gene rtxA. In order to investigate the gene functions of the secretion system, mutants of rtxB and rtxC were constructed respectively, resulting in VABM and VACM. The two mutants showed no significant change in exopolysaccharide production, whereas protease activity was decreased by 71% (VABM) and 58% (VACM),indicating that the RTX secretion system probably take part in the cross-membrane transport of protease, but not of exopolysaccharide.
Keywords: Vibrio anguillarum, secretion system, RTX cluster, mutant strain
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No.1575115604711926****
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