Detection of Transposon Mutants That Cannot Survive in Macrophages and Mice.

An all pairs experiment design type is where all labeled extracts are compared to every other labeled extract. We have adapted a microarray-based transposon tracking strategy for use with a Salmonella enterica serovar Typhimurium cDNA microarray in order to identify genes important for survival and replication in RAW 264.7 mouse macrophage-like cells or in the spleens of BALB/cJ mice. A 50,000-CFU transposon library of S. enterica serovar Typhimurium strain SL1344 was serially passaged in cultured macrophages or intraperitoneally inoculated into BALB/cJ mice. The bacterial genomic DNA was isolated and processed for analysis on the microarray. The novel application of this approach to identify mutants unable to survive in cultured cells resulted in the identification of components of Salmonella pathogenicity island 2 (SPI2), which is known to be critical for intracellular survival and replication. Keywords: all_pairs Computed

全配对实验设计（all pairs experiment design）指将所有标记提取物与其余每一份标记提取物进行比对的实验类型。本研究将基于微阵列的转座子追踪策略优化适配于肠炎沙门氏菌鼠伤寒血清型（Salmonella enterica serovar Typhimurium）的cDNA微阵列，以筛选可在RAW 264.7小鼠巨噬细胞样细胞或BALB/cJ小鼠脾脏中存活与增殖的关键基因。我们将该菌株SL1344的5万菌落形成单位（CFU）转座子文库，于培养巨噬细胞中连续传代，或通过腹腔接种至BALB/cJ小鼠体内。随后提取细菌基因组DNA并进行处理，以用于微阵列分析。将该方法创新性应用于筛选无法在培养细胞中存活的突变株，成功鉴定出沙门氏菌致病岛2（SPI2）的相关组分，而该致病岛已被证实对胞内存活与增殖至关重要。关键词：全配对实验设计（all_pairs Computed）