用地高辛配基标记DNA探针检测人类基因组单拷贝序列  

DETECTION OF HUMAN GENOMIC SINGLE COPY GENE SEQUENCE WITH DIGOXIGENIN-LABELED DNA PROBE

作　　者：蒋三亮 张思仲[1] 

机构地区：[1]华西医科大学医学遗传研究室

出　　处：《遗传与疾病》1991年第1期4-6,61+65,共3页

基　　金：国家自然科学基金;计划生育委员会基金

摘　　要：以地高辛配基标记DNA探针,通过Southern印迹杂交进行了检测人类单拷贝序列的可行性研究,同时比较了地高辛配基和生物素标记的敏感性。结果表明:地高辛配基系统在打点检测条上可检出40fg低限探针DNA,在Southern印迹杂交中可检出45.5fg低限λDNA;而生物素系统相应检测低限为1.6pg和450fg以3μg人基因组DNA进行Southern杂交,地高辛配基系统可稳定检出人基因组单拷贝序列,而生物素系统未获成功。本实验还将地高辛配基标记方法初步用于性反转患者的基因诊断。A digoxigenin-labeled DNA probe pDP 1007 was used to detect human genomic single copy gene sequence by Southern hybridization and its sensitivity was compared with biotinylated probe, The results showed that, with the digoxigenin labeling system, a minimum of 40fg probe was detectable in filter, and a minimum of 45.5fg Lambda DNA could be detected by Southern hybridization, while with the biotin system, the corresponding amounts were 1.6pg and 450fg.With the digoxigenin-labeled probe pDP1007, human genomic single copy gene sequence was detectable by Southern hybridezation, but the biotin system failed to do so. Digoxienin-labeled DNA probe was also successfully used to analyze the molecular pathology of two patients with sex reverse syndrome.

关 键 词：地高辛配基 单拷贝基因 DNA探针 

分 类 号：R394.8[医药卫生—医学遗传学]