浅表播散型汗孔角化症致病候选基因的突变筛查  

Mutation screening of candidate genes causing disseminated superficial porokeratosis

作　　者：刘晨帆[1] 王震英[2] 宋怀东[3] 潘春明[3] 张莉[2] 

机构地区：[1]济南市传染病医院传染病科,济南250021 [2]山东大学附属省立医院皮肤性病科,济南250021 [3]上海交通大学附属瑞金医院国家基因组学重点实验室,上海200025

出　　处：《山东大学学报（医学版）》2013年第1期93-97,共5页Journal of Shandong University:Health Sciences

基　　金：山东省科委基金(2002BB1DBA4)

摘　　要：目的对已定位的浅表播撒型汗孔角化症(DSP)致病区段的候选基因进行大规模突变筛查及突变分析,以识别该病的致病基因。方法收集家系成员的临床资料及血液样本,抽提外周血基因组DNA和正常人皮肤组织RNA。利用皮肤组织RNA对候选基因进行RT-PCR,确定在皮肤中表达的基因,根据基因组序列设计PCR引物,用家系样本的DNA做模板,进行PCR扩增,对扩增产物进行纯化、测序,再采用生物信息学方法处理测序结果,寻找突变位点,识别致病基因。结果发现了D12s1671～D12s1605之间的24个基因在皮肤中表达。共筛查了56个基因,发现家系4所有患者中存在2个SNP,即:UHRF1BP1L基因c.4242T>G错义突变导致1414位密码子由苯丙氨酸转化为亮氨酸和ANO4基因c.2487G>A突变导致829位密码子发生了同义突变。结论发现了家系4所有患者中存在的2个SNP,排除了56个候选基因是DSP致病基因的可能性。Objective To identify the candidate genes causing disseminated superficial porokeratosis （DSP） by large- scale mutation screening and analysis of the located segments. Methods We collected the pedigree members＇ clinical data and blood samples. Afterwards the genomic DNA of peripheral blood and the RNA of the skin tissue from healthy persons were extracted. First of all, we processed RT-PCR with RNA by classical method. Then, we designed PCR primers and selected the genes expressed in the skin to performe PCR amplification with genomic DNA of individuals of the pedigree. Afterwards, the PCR products were purified and sequenced with 2 μL purified product. The mutation a- nalysis was performed for the sequence in order to find the disease gene causing DSP. Results We found 24 genes were expressed in the candidate region between D12s1671 to D12s1605 by RT-PCR. We screened 56 candidate genes. A c. 4242T 〉 G nonsynonymous mutation in UHRF1BP1L gene that resulted in a substitution phenylalanine for leucine at codon 1414 （ p. F1414L） and a c. 2487G 〉 A synonymous mutation at the codon 829 in ANO4 gene, were found in all of the patients in the DSP family. Conclusion 2 SNPs are found in all of the patients in the DSP family and 56 can- didate genes are excluded.

关 键 词：汗孔角化症 大规模突变筛查 致病基因 单核苷酸多态性 聚合酶链反应 逆转录聚合酶链反应 

分 类 号：R751[医药卫生—皮肤病学与性病学]