直接杂交法检测基因组的单拷贝基因

本文叙述一种用于基因组限制性图谱研究的Southern印迹术的改良方法。正常和地中海贫血DNA经限制性内切核酸酶完全消化及凝胶电泳分离后,进行干印,变性与中和处理,再和相应的珠蛋白基因探针杂交。修改过的凝胶杂交法具有简便、经济和省时等优点,可对单拷贝基因进行灵敏、可靠的检测,因此能用于遗传病的产前诊断和DNA多态性的分析。作者还就此法与现存的探针制备系统合用的可行性作了扼要讨论。

THE DETECTION OF SINGLE COPY GENES IN GENOME BY DIRECT HYBRIDIZATION TECHNIQUE

This paper describes an alternative of Southern blotting technique for the genomic restriction mapping studies.The normal and thalassemic DNAs are completely digested with restriction endonucleases and electrophoresed on agarose gel.The resulting gel is then dry-blotted,denatured,neutralized,and hybridized with appropriate globin genespecific probes.The modified direct gel hybridization procedure has the advantage of simplicity,inexpensiveness,and time-saving.It can provide sensitive and reliable detection for single copy genes thereby being applicable to the prenatal diagnosis of genetic diseases and the analysis of DNA polymorphisms.The potential combination of this method with exsisting probe-making systems has also been discussed.