副溶血性弧菌全基因组DNA芯片的研制和质量评价

Development and evaluation of whole-genome DNA microarray of Vibrio parahaemolyticus

【目的】研制副溶血性弧菌全基因组芯片,建立芯片杂交方法,并对芯片质量进行评价。【方法】利用副溶血性弧菌全基因组序列,挑选出4770条基因,PCR扩增各基因并将PCR产物纯化,点样制备芯片;设计了两个质控杂交组合,采用双色荧光杂交策略,对芯片质量进行评价;PCR方法验证部分芯片结果。【结果】芯片杂交与理论预期结果以及PCR验证结果完全一致。【结论】成功的研制了一批质量良好的副溶血性弧菌全基因组DNA芯片,并建立了基于DNA芯片的副溶血性弧菌比较基因组学技术平台,建立了一套系统的芯片数据分析的标准方法。

Objective We developed and evaluated a whole-genome DNA microarray of Vibrio parahaemolyticus.MethodsBased on the genomic sequences of V.parahaemolyticus,we chose a total number of 4770 genes,amplified them by PCR with specific primers,purified the PCR products and printed them onto glass slides.We performed two sets of hybridizations by the method of two-fluorescence comparative hybridization to evaluate the microarray quality,followed by PCR method to validate parts of microarray results.Results Microarray hybridization results were completely consistent with theory expectations and PCR verification results.ConclusionWe successfully developed a batch of good quality whole-genome DNA microarrays of V.parahaemolyticus,built up a method of microarray-based comparative genomic hybridization of V.parahaemolyticus and a set of microarray data analysis standard method.