猪链球菌2型全基因组DNA芯片的研制及基因表达谱技术平台的建立  

Development of Whole-Genome DNA Microarray of Streptococcus suis Serotype 2 and the Platform of Gene Expression Profiling

作　　者：曾小涛[1,2] 袁媛[2] 郑玉玲[2] 周冬生[2] 姜永强[2] 陈福生[1] 

机构地区：[1]华中农业大学食品科技学院,湖北武汉430070 [2]军事医学科学院微生物流行病研究所,病原微生物生物安全国家重点实验室,北京100071

出　　处：《生物技术通讯》2009年第5期658-661,共4页Letters in Biotechnology

基　　金：国家自然科学基金(30600023)

摘　　要：目的:研制猪链球菌2型(SS2)全基因组DNA芯片,建立SS2基因表达谱技术平台。方法:利用SS2全基因组序列,挑选出2194条基因,经PCR扩增出2156条基因并将产物纯化,点样制备芯片;将芯片用于表达谱研究,采用实时定量PCR验证表达谱结果,对芯片进行可靠性分析。结果:芯片杂交数据与实时定量PCR验证显示了较高的相关性,二者相关系数r=0.87。结论:研制了一批SS2全基因组DNA芯片,并建立了基于DNA芯片的表达谱技术平台。Objective： To develop a whole-genome DNA microarray of Streptococcus suis serotype 2（SS2） and build up a method of gene expression profiling. Methods： Based on the genomic sequences of SS2, a total number of 2 194 genes were chosen, 2 156 of those were successfully amplified by PCR and purified, the PCR products were finally printed onto glass slides; the microarray were applied to gene expression profiling and the results were compared and validated with quantitative real time-PCR（qRT-PCR）. Results： Microarray hybridization result was mainly consistent with theory expectations and PCR verification results. The comparison of hybridization and qRT-PCR results showed a positive correlation（r= 0.87）. Conclusion： We developed a batch of whole-genome DNA microarray of SS2 with good quality, established a platform of microarray-based gene expression profiling of SS2, and built up a set of microarray data analysis standard methods.

关 键 词：猪链球菌2型 DNA芯片 芯片杂交 基因表达谱 

分 类 号：Q789[生物学—分子生物学]