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首页> 外文期刊>Infection and immunity >Sequence Analysis of TnphoA Insertion Sites in Vibrio cholerae Mutants Defective in Rugose Polysaccharide Production
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Sequence Analysis of TnphoA Insertion Sites in Vibrio cholerae Mutants Defective in Rugose Polysaccharide Production

机译：霍乱弧菌突变体中无糖多糖生产中TnphoA插入位点的序列分析

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Vibrio cholerae can switch from a smooth to a wrinkled or rugose colony phenotype characterized by the secretion of a polysaccharide that enables the bacteria to survive harsh environmental conditions. In order to understand the genetic basis of rugosity, we isolated TnphoA-induced stable, smooth mutants of two O1 El Tor rugose strains and mapped the insertion sites in several of the mutants using a modified Y-adapter PCR technique. One of the TnphoA insertions was mapped to the first gene of thevps region that was previously shown to encode the rugose polysaccharide biosynthesis cluster. Three insertions were mapped to a previously unknown hlyA-like gene, also in thevps region. Five other insertions were found in loci unlinked to the vps region: (i) in the epsDgene (encodes the “secretin” of the extracellular protein secretion apparatus), (ii) in a hydG-like gene (encodes a ?54-dependent transcriptional activator similar to HydG involved in labile hydrogenase production in Escherichia coli, (iii) in a gene encoding malic acid transport protein upstream of a gene similar to yeiE of E. coli(encodes a protein with similarities to LysR-type transcriptional activators), (iv) in dxr (encodes 1-deoxy-d-xylulose 5-phosphate reductoisomerase), and (v) in the intergenic region of lpd and odp (encode enzymes involved in the pyruvate dehydrogenase complex formation). These data suggest the involvement of a complex regulatory network in rugose polysaccharide production and highlight the general utility of the Y-adapter PCR technique described here for rapid mapping of transposon insertion sites.
机译：霍乱弧菌可以从光滑的表型转变为皱纹的或皱纹的菌落表型，其特征是多糖的分泌使细菌能够在恶劣的环境条件下生存。为了了解皱纹的遗传基础，我们分离了Tn phoA 诱导的两个O1 El Tor皱纹菌株的稳定，平滑突变体，并使用修饰的Y-适配器绘制了多个突变体中的插入位点PCR技术。 Tn phoA 插入片段中的一个被定位到 vps 区域的第一个基因，该基因先前已被证明编码皱纹多糖生物合成簇。在 vps 区域中，三个插入被定位到以前未知的 hlyA 样基因。在与 vps 区域不相关的基因座中发现了另外五个插入：（i）在 epsD 基因中（编码细胞外蛋白分泌装置的“分泌素”），（ii ）中的一个类似 hydG 的基因（编码一个与HydG类似的？ 54 依赖性转录激活因子，参与了大肠杆菌中不稳定的氢化酶的产生，（ iii）在与大肠杆菌的 yeiE 类似的基因上游编码苹果酸转运蛋白的基因中（编码与LysR型转录激活因子相似的蛋白质），（ iv）在 dxr 中（编码1-脱氧-d-木酮糖5-磷酸还原异构酶），以及（v）在 lpd 和 odp的基因间区域em>（编码参与丙酮酸脱氢酶复合物形成的酶），这些数据表明复杂的调节网络参与了皱纹多糖的生产，并突出了此处描述的Y适配器PCR技术在转座子快速作图中的通用性。锯齿部位。

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