变性梯度凝胶电泳检测到四例新的FⅨ基因突变  

Four novel point mutations of factor Ⅸ gene detected by denaturing gradient gel electrophoresis

作　　者：王泳[1,2] 李震宇[1,2] 万海英[1,2] 程大卫 阮长耿[1,2] 

机构地区：[1]苏州医学院附属第一医院 [2]江苏省血液研究所血栓与止血研究室

出　　处：《中华血液学杂志》1998年第3期125-128,共4页Chinese Journal of Hematology

摘　　要：目的：研究凝血因子Ⅸ的基因突变。方法：应用聚合酶链反应（ＰＣＲ）和变性梯度凝胶电泳（ＤＧＧＥ）筛选技术，分析了１２例血友病Ｂ患者ＦⅨ基因的全部８个外显子，剪接区及３′，５′端部分非编码区，并对ＤＧＧＥ行为异常片段进行双脱氧链终止法ＤＮＡ序列分析。结果：发现４例新的基因突变，分别位于ＦⅨ基因第１号外显子Ｔ（ＴＧＴ）１１１Ｇ（ＧＧＴ），导致Ｃｙｓ－１９Ｇｌｙ；第３号内含子剪接位点Ｃ１０３８０Ｔ和第８号外显子Ａ（ＴＡＣ）３０９１８Ｇ（ＴＧＣ），导致Ｔｙｒ２６６Ｃｙｓ；第８号外显子Ａ（ＡＣＧ）３１００７Ｃ（ＣＣＧ），导致Ｔｈｒ２９９Ｐｒｏ。结论：阐明血友病Ｂ的点突变有助于进一步了解ＦⅨ蛋白各功能域的精细功能，并为血友病Ｂ家系携带者检测和产前诊断提供了一个重要的遗传信息。Objective:To identify factor Ⅸ gene mutations in patients with hemophilia B.Methods:The coding regions,splicing junction sites and part of the 5′ and 3′ flanking regions of factor Ⅸ gene were screened by polymerase chain reaction(PCR) and denaturing gradient gel electrophoresis(DGGE).Results:Four amplified fragments showed abnormal electrophoresis patterns,and sequencing of them demonstrated four novel point mutations including a T to G transition at nucleotide 10380 resulting in intron 3 covering the splicing site ,a A to G transition at nucleotide 30918 resulting in a substitution of Cys for Tyr at codon 266 in exon 8,and a A to C transition at nucleotide 31007 resulting in a substitution of Pro for Thr at codon 299 in exon 8.Conclusion:Detection of mutations by PCR,DGGE and sequencing was useful not only for understanding the structure function relationship of FⅨ,but also for the carrier detection and prenatal diagnosis.

关 键 词：血友病B FⅨ基因 基因突变 DGGE 

分 类 号：R554.102[医药卫生—血液循环系统疾病]