铜绿假单胞杆菌QS相关基因差异表达的研究

选取100个与铜绿假单胞杆菌（Pseudomonas aeruginosa）群感效应（quorum-sensing，QS）相关的基因，克隆这些基因片段于pMD-18T载体，测序鉴定，点样制备cDNA基因芯片。制备cy3-dCTP／cy5-dCTP标记的探针，与芯片杂交。初步研究了处于不同生长期的铜绿假单胞杆菌基因的表达差异。指数中期和平台初期相比，有9个QS基因表达量最著增加，有6个基因表达量显著下降。利用芯片做针对铜绿菌假单胞杆菌药物的筛选：妥布霉素（Tobramycin）给药后细菌基因发生差异表达。证明了该cDNA芯片用于药物筛选的可行性。在国内首次研制开发了QS相关基因的cDNA芯片。应用基因芯片技术建立的铜绿假单胞杆菌QS相关基因研究平台，为找到能较好抑制铜绿假单胞杆菌正常生长的药物研究提出新的解决方法。

Expression Diversity of Quorum-sensing-Related Genes in Pseudomonas aeruginosa

One hundred Pseudomonns neruginosa quorum-sensing-related genes were selected and their primers were synthesized. The fragments of specific sequences which are related QS genes were amplified by PCR. These verified sequences were inserted into the vector pMD-18T for sequencing. These DNA fragments were dotted onto glass slides to make eDNA microarray, tlybridization was performed with cy3/cy5-dCTP labeled probes. The scanning data of early stationary phase and mid-logarithmic phase indicated that 9 genes were up-regulated and 6 genes were down regulated. Undergoing the different medicines, we took tobramycin as an example to compare the expression diversity. The results confirm that the QS eDNA chip is useful, and may contribute to better understand the mechanism of quorum-sensing, and can help us find the new targets for restraining the growth of Pseudomonas aeruginosa.