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目的建立一种应用鼠疫耶尔森菌全基因组芯片研究黄连对鼠疫耶尔森菌抑制作用分子机制的方法。方法应用液体稀释法测定黄连对鼠疫耶尔森菌的最小抑菌浓度(MIC);鼠疫耶尔森菌全基因组芯片包含4 005条鼠疫耶尔森菌基因。基因芯片表达谱实验中,黄连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;逆转录合成cDNA;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SAM软件处理结果。结果获得了黄连作用鼠疫耶尔森菌的基因芯片表达谱。结论应用鼠疫耶尔森菌全基因组DNA芯片可以进行黄连抑制鼠疫耶尔森菌的分子作用机制研究。
OBJECTIVE: To establish a method for the molecular mechanism of the inhibition of Coptis chinensis Yersinia pestis using Yersinia pestis genome-wide microarray. Methods The minimum inhibitory concentration (MIC) of Coptis chinensis to Yersinia pestis was determined by liquid dilution method. The whole genome of Yersinia pestis contains 4 005 Yersinia pestis genes. Gene chip expression profiling experiments, Coptis Chinensis Yersinia pestis concentration of 10 × MIC, the role of time was 30 min; extraction and purification of Yersinia pestis total RNA; reverse transcribed cDNA; Cy3, Cy5 dye labeling; And Yersinia pestis whole genome chip hybridization; expression profiling obtained by chip scanner. Apply SAM software to process the results. Results The gene chip expression pattern of Yersinia pestis was obtained. Conclusion The molecular mechanism of Coptis chinensis inhibiting Yersinia pestis can be studied using Yersinia pestis whole genome DNA chip.