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

Abstract:

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 2μL purified product. The mutation analysis 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 candidate genes are excluded.

Key words: Porokeratosis; Cosmical mutation screening; Disease gene; Single nucleotide polymorphism； Polymerase chain reaction； Reverse transcription polymerase chain reaction