CAJ | 학술논문

目的：检测一先天性厚甲症（PC）家系的致病基因KRT6a、KRT6b、KRT16、KRT17，以期找到其可能的致病突变。方法常规收集该家系成员外周静脉血，同时采集同地区100名健康自愿者外周血作为正常对照。分别提取DNA，运用聚合酶链反应(PCR)扩增基因KRT6a、KRT6b、KRT16、KRT17的全部外显子及其侧翼内含子序列，产物纯化后直接行DNA测序，比对分析其基因突变位点和类型。结果 PC患者KRT6a基因第1号外显子存在错义突变c.521T>C（p.Phe174Ser），而正常家系成员和正常对照组均无此突变，KRT6b、KRT16和KRT17基因也未发现致病突变。结论该PC家系的患者存在一个错义突变——KRT6a基因第1号外显子c.521T>C（p.Phe174Ser）。该突变是导致PC发病的分子基础。
목적：검측일선천성후갑증（PC）가계적치병기인KRT6a、KRT6b、KRT16、KRT17，이기조도기가능적치병돌변。방법상규수집해가계성원외주정맥혈，동시채집동지구100명건강자원자외주혈작위정상대조。분별제취DNA，운용취합매련반응(PCR)확증기인KRT6a、KRT6b、KRT16、KRT17적전부외현자급기측익내함자서렬，산물순화후직접행DNA측서，비대분석기기인돌변위점화류형。결과 PC환자KRT6a기인제1호외현자존재착의돌변c.521T>C（p.Phe174Ser），이정상가계성원화정상대조조균무차돌변，KRT6b、KRT16화KRT17기인야미발현치병돌변。결론해PC가계적환자존재일개착의돌변——KRT6a기인제1호외현자c.521T>C（p.Phe174Ser）。해돌변시도치PC발병적분자기출。
ObjectiveTo test the four causative keratin genes, namely, KRT6a, KRT6b, KRT16 and KRT17, in a Chinese pedigree with pachyonychia congenita (PC), so as to identify the possible pathogenic mutation which lead to the occurrence of the disease. MethodsPeripheral venous blood from every members of the Chinese PC family and a panel of 100 unaffected control individuals matched for the geographical location were routinely collected. Genomic DNA was extracted from these blood samples. All the coding exons and their lfanking intronic sequences of KRT6a,KRT6b, KRT16 and KRT17 genes were ampliifed by polymerase chain reaction(PCR), then these products were puriifed for direct DNA sequencing. At last, the gene mutation sites and types were identiifed by comparative analysis. ResultsA missense mutation, c.521T>C (p.Phe174Ser) in 1 exon of KRT6a gene was identiifed in two PC patients, which was not found in the healthy members and 100 normal controls. Meanwhile, no other mutation was found in KRT6b, KRT16 and KRT17 genes.ConclusionsA missense mutation, c.521T>C (p.Phe174Ser), in 1 exon of KRT6a was identiifed in the pedigree with PC which we investigated, it has been demonstrated to be the molecular basis of PC pathogenesis.