Expression Analysis of WR99210-treated Mycobacterium tuberculosis H37Rv. Mycobacterium tuberculosis H37Rv

Investigation of whole genome gene expression level changes in Mycobacterium tuberculosis treated with the DHFR inhibitor WR99210, compared to untreated cells. The antimycobacterial properties of WR99210 are further described in Gerum, A., Ulmer, J., Jacobus, D., Jensen, N., Sherman, D., and C. Sibley. 2002. Novel Saccharomyces cerevisiae screen identifies WR99210 analogues that inhibit Mycobacterium tuberculosis dihydrofolate reductase. Antimicrob Agents Chemother 46(11):3362-3369 [PMID:12384337] Overall design: A nine chip experiment in which total RNA was collected from H37Rv after treatment with 100 uM WR99210 for 0, 4, and 24 hours. Three separate cultures/biological replicates were interrogated for both treated and untreated cultures. Labeled cDNA was hybridized to NimbleGen spotted oligo tiling arrays covering both strands of the M. tuberculosis genome at ~200 bp intervals between 60-mer probes.

本研究探究经二氢叶酸还原酶（DHFR）抑制剂WR99210处理的结核分枝杆菌（Mycobacterium tuberculosis）与未处理细胞的全基因组基因表达水平变化差异。WR99210的抗分枝杆菌特性已在以下文献中详细阐释：Gerum A、Ulmer J、Jacobus D、Jensen N、Sherman D与Sibley C于2002年发表的研究。该研究通过酿酒酵母（Saccharomyces cerevisiae）筛选体系，发现了可抑制结核分枝杆菌二氢叶酸还原酶的WR99210类似物，刊载于《Antimicrob Agents Chemother》第46卷第11期，页码范围3362-3369[PubMed编号：12384337]。实验设计概述：本实验共使用9张芯片，从经100 μM WR99210分别处理0、4、24小时的H37Rv菌株中提取总RNA。处理组与未处理组均设置3组独立培养物作为生物学重复，并对每组重复进行检测分析。将标记后的cDNA与NimbleGen斑点寡核苷酸平铺芯片进行杂交，该芯片覆盖结核分枝杆菌基因组的两条链，探针间距约200 bp，探针长度为60聚体。