Detection of bacterial virulence genes by subtractive hybridization:: Identification of capsular polysaccharide of Burkholderia pseudomallei as a major virulence determinant
Detection of bacterial virulence genes by subtractive hybridization:: Identification of capsular polysaccharide of Burkholderia pseudomallei as a major virulence determinant
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DOI：
10.1128/iai.69.1.34-44.2001
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发表时间：
2001-01-01
影响因子：
3.1
通讯作者：
Woods, DE
中科院分区：
文献类型：
--
作者：
Reckseidler, SL;DeShazer, D;Woods, DE
类鼻疽伯克霍尔德氏菌是类鼻疽病的病原体，可引起人类和动物的广泛疾病，特别是在东南亚和北方澳大利亚，在那里它是地方病。泰国伯克霍尔德菌是一种与B密切相关的非致病性环境生物。假鼻疽在这两个物种之间进行消减杂交，以鉴定编码B中毒力决定簇的基因。假鼻疽消减文库的筛选揭示了B特有的富含A-T的DNA序列。假鼻疽，表明它们可能是通过水平转移获得的。其中一个消减克隆pDD 1015编码的蛋白质与铜绿假单胞菌的糖基转移酶具有同源性。该基因在野生型B中被插入失活。产生SR 1015。酶联免疫吸附试验和免疫电镜结果表明，失活的基因参与了一种主要表面多糖的产生。野生型B的半数致死量（LD 50）。假鼻疽是
Burkholderia pseudomallei, the etiologic agent of melioidosis, is responsible for a broad spectrum of illnesses in humans and animals particularly in Southeast Asia and northern Australia, where it is endemic. Burkholderia thailandensis is a nonpathogenic environmental organism closely related to B. pseudomallei. Subtractive hybridization was carried out between these two species to identify genes encoding virulence determinants in B. pseudomallei. Screening of the subtraction library revealed A-T-rich DNA sequences unique to B. pseudomallei, suggesting they may have been acquired by horizontal transfer. One of the subtraction clones, pDD1015, encoded a protein with homology to a glycosyltransferase from Pseudomonas aeruginosa. This gene was insertionally inactivated in wild-type B. pseudomallei to create SR1015. It was determined by enzyme-linked immunosorbent assay and immunoelectron microscopy that the inactivated gene was involved in the production of a major surface polysaccharide. The 50% lethal dose (LD50) for wild-type B. pseudomallei is