生物素标记DNA探针测定人性别

Biotinylated Y Chromosome Specific Probe for Human Sexing

白细胞或胎儿绒毛组织染色体DNA经限制酶Hae (?)消化后,用光生物素(photo-biotin)标记人Y染色体特异的3.4 kb DNA片段作探针,进行Southern印迹杂交,并以生物素结合蛋白-碱性磷酸酶体系显示生物素探针杂交信号。结果表明,0.1μg男性DNA可显示清晰的信号,而5μg女性DNA则无信号出现;平行试验证明生物素探针测定人性别的结果与32P标记探针完全一致。提示生物素探针可用于X连锁遗传病、性异常,运动员体检,法医学及异性别骨髓移植方面的性别分析。

Human chromosome DNA from WBC or fetus chorion samples were digested with Hae 333 and hybridized with biotinylated Y chromosome specific probe by Southern blotting, and hybridization signals were developed by the ABC (Avidin-biotin-alkaline phosphatase complex) system. The hybridization signal for 0.1μg of male DNA could be detected clearly, while the signal for even 5μg of female DNA could not. Parallel tests showed that the sexing results using 32P-labeled and biotinylated Y probe were identical. This suggests that the biotinylated Y probe can be applied to the determination of X-linked genetic diseases and sex abnormality, forensic analysis, sex determination of sportsmen and women, heterosexual transplanation of bone marrow, etc. It could become a convenient means for genetic diagnosis.