VIBRIO GENE EXPRESSION DURING INFECTION

描述（改编自申请人摘要）： 感染小肠粘膜表面的霍乱弧菌， 这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过程中表达，以及 调节蛋白和控制其调节的宿主信号， 基本的 构建了一个新的基因报告系统， 鉴定霍乱弧菌的体内诱导基因，并已用于 鉴定vie（体内表达的霍乱弧菌）操纵子。 Vie预测 编码三个双组分信号转导蛋白，包括 传感器和两个响应调节器，第一个这样的系统描述在V。 胆汁。 在这些系统中，VIE的操纵子结构和 它在感染期间的唯一表达。 基于与其他 系统，假设Vie蛋白通过感知 宿主肠道的环境条件，并通过调节 其他基因 因此，对vie的描述提供了一个独特的机会， 既可以探测宿主信号， 控制霍乱弧菌的生长和致病性， vie调控基因的研究。 本研究提案的目标 是描述vie操纵子的特征， vie调控基因 突变将在vie和 vie-regulated基因，以及每一个对致病性的影响将被 测定了 将确定Vie蛋白的亚细胞位置 用免疫化学的方法。 时空表达模式 vie操纵子和vie调控基因的数量将在 感染 这项工作可能有助于粘膜疫苗的开发， 涉及用于抗粘膜病原体治疗的新靶点的鉴定。

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.