肝豆状核变性分子生物学研究

【目的】探讨中国人肝豆状核变性(WD)的分子发病机制和基因诊断的方法。【方法】应用基因工程技术对WD进行了10年的分子生物学研究。【结果】①WD的基因定位研究通过RFLP及微卫星多态性分析，应用两位点及多位点连锁软件，建立了中国人WD基因在D

The Molecular Biological Study of Hepatolenticular Degeneration

【Objective】 To investigate the molecular basis of hepatolenticular degeneration (Wilson disease, WD) and to attempt to construct the feasibility of gene diagnosis in the disease. 【Methods】 We have performed the molecular biological study on this disease for 10 years by molecular geneti c techniques. 【Results】 ①Location of WD gene in Ch inese: Using pairwise linkage analysis and multipoint linkage analysis method, w e constructed a genetic map of DNA markers within D13q14.2-3 which refined the location of WD gene by restriction fragment length polymorphism(RFL P) and microsatellite polymorphism analysis; ②Screen for mutations of WD gene in Chinese people: we detected the structure of 21 exons of WD ge ne in 45 patients from 39 pedigrees by PCR-SSCP(Single strand conformation poly morphism) and PCR-DNA sequencing technology, found a new mutation in exon 5 and nuclcotide sequence analysis showed it is a T insertion. We also conformed the Arg778Leu in exon 8, the highest frequence mutation point in Chinese people, wit h mutation rate 22.8% in total;③Carrier detection and presymptomatic diagnosi s of WD: Based on DNA recombination technology, we peformed successfully the gen e diagnosis in all individuals of 79 families with WD and built up a helpful spe cific enzyme cut method (PCR-Msp1) to detect the carrier and presympomatic patients in Chinese pe ople with WD. 【Conclusion】 These results showed that the location of WD gene within D13q14.2-3 is the same in Chinese as in Caucasians, but the g ene high m utation point,the gene diagnosis method and its pathogenesis are markly different.