Screening of CYP21 gene P459H mutation by PCR-ACRS / 中国病理生理杂志

ABSTRACT

AIM: To investigate the frequency of P459H(CCC→CAC) in exon 10 of CYP21 gene among normal population.METHODS: The exons 3-10 of CYP21 gene were amplified with polymerase chain reaction(PCR).The PCR round products were digested by restriction enzyme Pst I to confirm that CYP21 gene was specifically amplified.PCR-based amplification-created restriction site(PCR-ACRS) was performed using the first round PCR products as template.After the second PCR products were digested by restriction enzyme Fsp I,10% polyacrylamide gel electrophoresis was used to screen the frequency of P459H in exon 10.RESULTS: The codon 459 in exon 10 of CYP21 gene was all CCC among 100 normal cases tested.CONCLUSION: P459H(CCC →CAC) of CYP21 gene might be a novel point mutation causing CAH.Furthermore,PCR-ACRS was a fast and safe method for gene mutation screening.