X连锁显性遗传性肾炎一家系相关基因排除定位分析.pdfVIP

1508 Journal of Chinese Physician, November,2004,Vol6,NolI x.连锁显性遗传性肾炎一家系相关基因排除定位分析 贺晓蕾，戴勇，王沙燕，黄瑞芳，石之磷，张阮章 (暨南大学医学院第二附属医院深圳市人民医院肾内科，广东 深圳 518020) 【摘要】目的 研究我国X连锁显性遗传性肾炎一家系(又名Alport综合征，Alport s syndrome, AS)编码W型胶原0链的 COIAA5基因突变特点。方法 提取35例现存家系成员基因组DNA，根据C01AA5基因中与外显子相郁的内含子序列设计引物，分 别作DNA聚合酶链式反应(PCR)，将该基因所有51个外显子扩增。以琼脂糖凝胶电泳鉴定产物，作单链构象多态性(SSCP)分析 检列PCR产物，对迁移率异常者进行DNA直接测序。结果 PCR - SSCP分析发现17个外显子PCR产物单链DNA迁移率异常，通 过DNA直接测序发现9个外显子可疑突变，但反向测序均未能证实。结论 该家系C014A5基因外显子未发生突变，其突变可能 发生在该基因内含子或其它致病基因。 〔关键词] Alport综合征;聚合酶钱反应;COL4A5基因 A Exclusive Mapping Analysis on the Gene Associated with X一】inked Dominant Inherited Nephritis in One Pedigree }lX 一sS r p4l.T{ , T4YVt SAa一ran，ft al. Medical Research Center, Shenzhen People Hospital, Sheuzhea 518020, China (Abstract]ObjectiveTo study the characteristics of mutations of gene C014A5 encoding type 1V collagen a 5 chain in one chinese pedi- gree with X一linked dominant inherited nephritis (Alport s syndrome, AS).MethodsGenomic DNA was extracted from 35 members of the pedi- gree of Alports syndrome. All of 51 exons of C014A5 gene were amplified by PCR with the primers synthesized according to the published intron se- quences of COIAA5.The PCR products were further analyzed by agarose gel electrophoresis and single strand conformation州ymorphism (SSCP). The PCR products showing polymorphism were directly sequenced. ResultsPCR一SSCP analysis showed that 17 PCR products had abnormal mobil- ity of single strand DNA. DNA sequencing analysis revealed 9 suspicious mutations. But these suspicious mutations were not be confirmed by inverse sequencing analysis. Conclusion The exon mutation of C014A5 gene of this pedigree did not be found, and the mutations of COIAA5 gene may lo- cate in the its introns. (Key words Alport s syndrome; Polymerase chain reaction; CO1AA5