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.
出处
《解放军医学杂志》
CAS
CSCD
北大核心
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
关键词
耶尔森氏菌
鼠疫
DNA芯片
比较基因组学
Yersinia pestis
whole-genome DNA microarray
comparative genomics
