Genetic tools for analyzing B. pseudomallei virulence

描述（由申请人提供）：拟议研究的目标是开发假芽孢杆菌遗传操作的新方法。待开发的工具应该能够更容易地创建靶向突变和多突变菌株，识别非转座子插入的未知突变，对基因组的任何区域进行互补分析，以及生成和定义具有杂交基因组的菌株。拟议的研究符合R21资助机制的探索性/开发性质，因为它旨在开发具有潜在广泛影响的程序，以实现项目公告（PA-03-080）的目标，例如鉴定假芽孢杆菌毒力决定因素并分析其功能，指定先天和获得性抗生素耐药机制，鉴定宿主识别的细菌抗原，以及创建活疫苗菌株。

DESCRIPTION (provided by applicant): The goal of the proposed research is to develop new methodologies for the genetic manipulation of B. pseudomallei. The tools to be developed should make it easier to create targeted mutations and multiple mutant strains, to identify unknown mutations not due to transposon insertion, to carry out complementation analysis for any region of the genome, and to generate and define strains with hybrid genomes. The proposed research is consistent with the exploratory/development nature of the R21 funding mechanism because it is designed to develop procedures with potentially broad impact on reaching the goals of the program announcement (PA-03-080), such as identifying B. pseudomallei virulence determinants and analyzing how they function, specifying innate and acquired antibiotic resistance mechanisms, identifying bacterial antigens recognized by the host, and creating live vaccine strains.