酶标DNA探针与Y染色体DNA的探测

 用辣根过氧化物酶标记DNA的技术,制备了酶标基因探针。研究了酶标过程和产物的电泳行为;用斑点杂交和southern印迹杂交探测了单链、双链DNA,灵敏度可达pg水平,以此酶标的Y染色体特异的DNA片段作探针,进行了DNA杂交的性别分析,证明该探针能清楚地区别两性基因组DNA,这对基因的研究和诊断有一定实用价值。

ENZYME-LABELED DNA PROBES AND THE DETECTION OF Y CHROMOSOME DNA

A study on enzyme-labeled probe for DNA hybridization was carried out. The DNA fragment or recombinant plasmid labeled with Horseradish peroxidase was used as probe to hybridize with single or double stranded DNA in dot or Southern blots. The technique was sensitive to DNA at pg level. Peroxdase-labeled Y-chromo-some specific DNA was used as a sex probe to examine the human WBC genomic DNAs from different sex. The result showed that the male DNAs could be clearly distinguished from the female DNAs by dot blots. It is, therefore, suggested that the enzyme-labeled DNA probe could be used in stead of radioactive probe for sex determination, as well as extensively applied to (the analysis of multicopy genes for diagnosis of certain diseases.