鼠疫耶尔森菌全基因组DNA芯片的研制及用于比较基因组学分析  

Development of whole-genome DNA microarray of Yersinia pestis and its validity in comparative genomic analysis

作　　者：周冬生[1] 韩延平[1] 戴二黑[1] 宋亚军[1] 包静月[2] 裴德翠[1] 李敏[3] 崔百忠[3] 张秀清[2] 童宗中[2] 王津[1] 郭兆彪[1] 祁芝珍[3] 金丽霞[3] 翟俊辉[1] 杜宗敏[1] 王效义[1] 汪建[2] 黄培堂[1] 杨瑞馥[1] 

机构地区：[1]北京军事医学科学院微生物流行病研究所,100071 [2]中国科学院北京基因组研究所 [3]青海省地方病预防控制所

出　　处：《解放军医学杂志》2004年第3期200-203,共4页Medical Journal of Chinese People's Liberation Army

基　　金：国家"8 63"基金 (编号 2 0 0 1AA2 2 30 61 );国家自然科学基金 (编号30 371 2 84)资助课题

摘　　要：目的　研制鼠疫耶尔森菌全基因组DNA芯片 ,并将其用于比较基因组分析。方法　挑选出 4 0 0 5条鼠疫耶尔森菌基因 ,PCR扩增各基因 ,以纯化的PCR产物点样制备芯片 ,采用双色荧光杂交策略 ,进行芯片比较基因组杂交。结果　设计了若干质控杂交组合 ,芯片杂交结果与全基因组测序结果完全一致。Objective To develop a whole-genome DNA microarray based on the genomic sequences of Y. pestis CO92 and 91001 and its use in comparative genomic analysis. Methods A total number of 4 005 genes of Y. pestis were amplified by PCR and printed onto glass slides in duplicate. Fluorescently labeled probes were prepared by marking genomic DNAs with random hexamers and Klenow. Labeled DNAs were hybridized with the microarrays by the method of two-fluorescence comparative hybridization. Three sets of two-fluorescence hybridizations were performed to examine the absence/presence of each gene. Results The results agreed with those derived from the in silico genomic comparison. Conclusion The results demonstrate that the microarry can be a useful tool for comparative genomic analysis of Y. pestis.

关 键 词：耶尔森氏菌 鼠疫 DNA芯片 比较基因组学 

分 类 号：R378.61[医药卫生—病原生物学]