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伯纳特柯克斯体基因编码一种类似传感器的蛋白质。

A Coxiella burnetii gene encodes a sensor-like protein.

作者信息

Mo Y Y, Mallavia L P

机构信息

Department of Microbiology, Washington State University, Pullman 99164-4233.

出版信息

Gene. 1994 Dec 30;151(1-2):185-90. doi: 10.1016/0378-1119(94)90654-8.

Abstract

Two-component regulatory systems play important roles in the adaptive responses of many bacteria to environmental changes. The sensor proteins of these systems are highly conserved near their C-termini. We exploited this feature to isolate a gene encoding a putative sensor component from the obligate intracellular rickettsial parasite Coxiella burnetii (Cb). Using degenerate primers and the polymerase chain reaction (PCR), we isolated a DNA fragment from a genomic library of Cb containing an open reading frame (ORF), sufficient to encode a 48-kDa protein. Sequence comparison revealed that the deduced protein shared high homology to members of the bacterial sensor protein family, particularly at three conserved regions of the C terminus. When the Cb sensor-like gene was cloned into a high-copy-number vector and introduced into an E. coli strain (phoM, phoR), the mutant expressed low levels of alkaline phosphatase activity, suggesting that the gene functioned as a sensor protein in E. coli. Consequently, we designated this gene qrsA (for Q fever agent regulatory sensor-like gene). Because two-component regulatory systems have been implicated in a variety of cellular processes, including virulence determinants in some pathogenic bacteria, the identification of qrsA in Cb may shed light on how the pathogen adapts to extracellular changes during infection, as it proliferates in the phagolysosome.

摘要

双组分调控系统在许多细菌对环境变化的适应性反应中发挥着重要作用。这些系统的传感蛋白在其C末端附近高度保守。我们利用这一特性从专性细胞内立克次氏体寄生虫伯氏考克斯体(Cb)中分离出一个编码假定传感成分的基因。使用简并引物和聚合酶链反应(PCR),我们从Cb的基因组文库中分离出一个DNA片段,该片段包含一个开放阅读框(ORF),足以编码一个48 kDa的蛋白质。序列比较显示,推导的蛋白质与细菌传感蛋白家族成员具有高度同源性,特别是在C末端的三个保守区域。当将Cb类传感基因克隆到高拷贝数载体中并导入大肠杆菌菌株(phoM,phoR)时,该突变体表达的碱性磷酸酶活性水平较低,这表明该基因在大肠杆菌中作为传感蛋白发挥作用。因此,我们将该基因命名为qrsA(用于Q热病原体调控传感样基因)。由于双组分调控系统涉及多种细胞过程,包括一些病原菌中的毒力决定因素,在Cb中鉴定qrsA可能有助于揭示该病原体在吞噬溶酶体中增殖时如何适应感染期间的细胞外变化。

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