摘要目的 研究基因组芯片检测大黄抑制鼠疫耶尔森菌分子机制的方法 .方法 使用的鼠疫耶尔森菌全基因组芯片包含4005条鼠疫耶尔森菌基囚.应用液体稀释法测定大黄对鼠疫耶尔森菌的最小抑菌浓度(MIC),基因芯片表达谱实验中,大黄作用鼠疫耶尔森菌的浓度为10×MIC,作用时间为30 min,提取并纯化鼠疫耶尔森菌总RNA,反转录合成cDNA,用Cy3,Cy5染料标记后,与鼠疫耶尔森菌全基因组芯片杂交,通过芯片扫描仪获得表达谱分析结果 .应用实时定量PCR技术对芯片结果 进行验证.结果 大黄对鼠疫耶尔森菌的MIC为0.02500 kg/L.获得了大黄作用鼠疫耶尔森菌的基因芯片表达谱.大黄作用鼠疫菌的明显差异表达基凶共有498个,上渊基闪358个,下凋基因140个.结论 应用鼠疫耶尔森菌全基因组DNA芯片可以进行大黄抑制鼠疫耶尔森菌的分子作用机制研究.

abstractsObjective To establish a method for studying molecular mechanism of Rhubarb inhibiting anti-Yersinia pesti based on DNA microarray.Methods A whole genome DN A microarray containing 4005 annotated genes of Yersiniapesti Was used.The minimal inhibitory concentration(MIC)of Rhubarb to Yersiniapestiwas determined by liquid dilution method.The gene expression profile of Yersinia pesti was performed after the exposure to Rhubarb at a concentration of 10×MIC for 30 minutes.The total RNA extracted and purified from Yersinia pesti Was reversely transfected to cDNA and labeled by Cy3-Cy5 dye.The labeled probes were hybridized to the microarray anti the results were obtained by a laser scanner and the microarray data was confirmed by real-time quantitative RT-PCR.Results The platform of the DNA microarray-based bacteria transcriptional profile was established.A total of 498 genes of Yersinia pesti changed significantly in response to Rhubarb.Among them.358 genes were up-regulated,140 down-reguated.Conclusions The whole genome DNA microarray can be used in the studying of molecular anti-Yersinia pesti mechanism of Rhubarb.