【摘要】 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。更多还原

【Abstract】 Objective To test the four causative keratin genes, namely, KRT6 a, KRT6 b, 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. Methods Peripheral 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 flanking intronic sequences of KRT6 a,KRT6b, KRT16 and KRT17 genes were amplified by polymerase chain reaction(PCR), then these products were purified for direct DNA sequencing. At last, the gene mutation sites and types were identified by comparative analysis. Results A missense mutation, c.521T>C(p.Phe174Ser) in 1 exon of KRT6 a gene was identified in two PC patients, which was not found in the healthy members and 100 normal controls. Meanwhile, no other mutation was found in KRT6 b, KRT16 and KRT17 genes. Conclusions A missense mutation, c.521T>C(p.Phe174Ser), in 1 exon of KRT6 a was identified in the pedigree with PC which we investigated, it has been demonstrated to be the molecular basis of PC pathogenesis.更多还原