摘要
目的对一个遗传性凝血因子Ⅴ缺乏症家系进行凝血因子Ⅴ(factorⅤ,FⅤ)基因突变的检测,探讨先天性凝血因子Ⅴ缺乏症的分子发病机理。方法PCR结合直接测序方法对先证者的FⅤ基因全部25个外显子及其旁侧序列进行分析,鉴别其中可能存在的基因变异。应用PCR产物反向测序法或PCR限制性内切酶分析技术对突变位点进行确证。随机选择100名健康体检者作为正常对照。结果先证者FⅤ基因存在两种突变,分别是第11外显子区的A1763C错义突变和位于第16内含子3′端剪接位点的G→T突变;家系分析表明这两个突变是双杂合子型,前者遗传自父亲,后者遗传自母亲。100名健康对照者均未发现A1763C错义突变。结论第11外显子A1763C错义突变和第16内含子3′端的剪接位点突变使先证者呈现双重杂合子型,可能是先证者FⅤ先天性缺乏的原因。
Objective To discover the mutations of human blood coagulation factor Ⅴ （FⅤ） gene in a Chinese lamily with eongenital factor Ⅴ deficiency, and to explore the molecular mechanism associated with the congenital factor Ⅴ deficiency. Methods PCR and DNA sequencing were used to look for the FⅤ gene mutations in the proband. And the novel mutation were testified by PCR restriction fragment length polymorphism technique or reverse DNA sequencing. One handred healthy volunteers were chosen as controls at random. Results Two novel mutations were discovered in the FⅤ gene of proband, which were the A1763C missense mutation in exon 11 and the splicing site mutation in the 3＇ terminal of intron 16 （G→T）. The pedigree analysis showed that the two mutations inherited from Iris parents respectively： the A1763C came from his father, and the G→T from his mother. The A1763C missense mutation in exon 11 was not found in each of 100 healthv volunteers. Conclusion The congenital deficiency of FⅤ in the proband might be caused by the A1763C missense mutation in exon 11 and the splicing site mutation in the 3＇ terminal of intron 16, which jointly caused the proband to be a double heterozygote.
出处
《中华医学遗传学杂志》
CAS
CSCD
北大核心
2006年第5期515-518,共4页
Chinese Journal of Medical Genetics
基金
广东省自然科学基金(06020907)
广东省人民医院科研基金(Y204051)~~
关键词
凝血因子Ⅴ
突变
聚合酶链反应
coagulation factor Ⅴ
mutation
polymerase chain reaction
