VIBRIO GENE EXPRESSION DURING INFECTION

描述(改编自申请人的摘要)：致病性 霍乱弧菌感染小肠粘膜表面，是 被认为是一个协调和多因素的过程。不幸的是， 了解哪些基因在感染期间表达，以及 控制其调控的调节蛋白和宿主信号，是 很初级的。构建了一种新的基因报告系统，用于 霍乱弧菌体内诱导基因的鉴定，并已用于 鉴定VIE(霍乱弧菌体内表达)操纵子。VIE是预测的 编码三个两组分的信号转导蛋白，包括一个 传感器和两个响应调节器，这是V。 霍乱弧菌。VIE在这类系统中是独一无二的，它的操纵子结构和 其在感染过程中的独有表达。基于对其他 系统，假设VIE蛋白的功能是通过感知 寄主肠道的环境条件及其调控反应 其他基因。因此，VIE的表征提供了一个独特的机会 既可以探测宿主信号，也可以探测细菌调节蛋白 控制霍乱弧菌的生长和致病性，并允许 VIE调控基因的研究。这项研究提案的目标是 是确定VIE操纵子的特征，并识别和表征 VIE调控的基因。突变将在VIE和VIE中构建 VIE调控的基因，每个基因对致病性的影响将是 量过了。VIE蛋白的亚细胞位置将被确定 使用免疫化学方法。空间和时间的表达模式 VIE操纵子和VIE调控基因的多少将在 感染。这项工作可能有助于粘膜疫苗的开发和 以确定针对粘膜病原体治疗的新靶点。

DESCRIPTION (Adapted from the applicant's abstract): The pathogenicity of Vibrio cholerae, which infects mucosal surfaces in the small intestine, is believed to be a coordinated and multifactorial process. Unfortunately, knowledge of which genes are expressed during infection, as well as the regulatory proteins and host signals that govern their regulation, is rudimentary. A novel gene reporter system has been constructed for identifying in vivo induced genes of V. cholerae, and has been used to identify the vie (V. cholerae in vivo expressed) operon. vie is predicted to encode three two-component signal transduction proteins, including a sensor and two response regulators, the first such system described in V. cholerae. vie is unique among such systems both in its operon structure and its exclusive expression during infection. Based on analogy to other systems, it is hypothesized that the Vie proteins function by sensing environmental conditions in the host intestine and responding by regulating other genes. Characterization of vie thus provides a unique opportunity both to probe the host signals and bacterial regulatory proteins that control V. cholerae growth and pathogenicity as well as to allow investigation of vie-regulated genes. The goals of this research proposal are to characterize the vie operon and to identify and characterize vie-regulated genes. Mutations will be constructed in both vie and vie-regulated genes, and the effects of each on pathogenicity will be measured. The subcellular locations of the Vie proteins will be determined using immunochemical methods. The spatial and temporal expression patterns of the vie operon and vie-regulated genes will be determined during infection. This work may contribute to development of mucosal vaccines and to identification of new targets for therapies against mucosal pathogens.