cDNA文库法制备HCV-1诊断芯片探针的应用性研究  

DNA Microarray Probe Preparation Us ing cDNA Fragments Library from HCV Genotype 1

作　　者：孙朝晖[1] 郑文岭[2] 张宝[1] 彭翼飞[1] 马文丽[1] 

机构地区：[1]第一军医大学基因工程研究所,中国广东广州510515 [2]广州军区广州总医院肿瘤分子生物学研究所,中国广东广州510010

出　　处：《生命科学研究》2004年第3期256-259,共4页Life Science Research

基　　金：国家自然科学基金资助项目(39880032);广州市重点科技攻关项目(99-Z-022-01)

摘　　要：探讨利用cDNA文库法制备HCV-1诊断基因芯片探针的可行性.用限制性内切酶Sau3AⅠ消化HCV1a及1b全长cDNA,所得的酶切片段72℃补平加A,AT克隆,PCR初步鉴定,并测序.结果显示:HCV两个亚型1a、1b的全长cDNA得到57个大小相对一致(200～1000bp)的片段,平均每个亚型约28个,PCR及序列分析表明,所扩增的片段均属于HCV-1的特异基因,可做为HCV-1诊断基因芯片探针.利用cDNA文库法收集片段是一种快速、简便制备芯片探针的实用方法.The technique of cDNA fragments libr ary construction in the preparation of HCV-1probes for clinical diagnostic microarray were studied.The full-length HCV cDNAs of distinct subtypes of 1a and 1b were digested with restriction endonuclease Sau3A I and the resulting fragments were cloned into the pMD18-T vectors.Positive clones were isola ted and identified by sequencing.A t otal of 57different fragments were obtained and sequences analysis showed that all the fragments ranging fr om 200to 1000bp were specific gene fragments of HCV genotype 1,which ca n be efficiently used in microarray p repapration.The method of preparing microarray probes by construction of cDNA fragments library was effective,quick and simple.

关 键 词：HCV 分子克隆 微阵列 分子探针 

分 类 号：R394.2[医药卫生—医学遗传学]