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目的对一先天性无虹膜症家系进行了致病基因PAX6的突变分析。方法PCR反应扩增PAX6基因的所有外显子,PCR产物进行SSCP(单链构象多态性)分析,通过患者与正常人带型的差异来确定突变发生的外显子,对有差异SSCP带型的PCR产物进行直接测序找到突变位点。PCR产物进一步亚克隆到pGEM-T载体,测序验证突变位点。结果发现基因突变为PAX6基因第2内含子和第3外显子之间的剪接识别位点“AG”中的碱基A的丢失(IVS2-2delA)。结沦PAX6基因5′端非编码区剪接突变能够引起先天性无虹膜症。
Objective To analyze the mutation of PAX6 gene in a pedigree of congenital absence of iris. Methods All the exons of PAX6 gene were amplified by PCR. PCR products were analyzed by single strand conformation polymorphism (SSCP). The differences of the genotypes between the patients and the controls were used to determine the exon. Type of PCR products were directly sequenced to find the mutation site. PCR products were further subcloned into pGEM-T vector and sequenced to verify the mutation sites. As a result, it was found that the gene mutation was a loss of base A (IVS2-2delA) in the splicing recognition site “AG” between the second intron and the third exon of the PAX6 gene. Knot degeneration PAX6 gene 5 ’non-coding region splicing mutations can cause congenital absence of iris syndrome.