巢式PCR-RFLP法检测Cystatin C基因多态性的实验研究

Determination of CST3 Gene Polymorphism by Nested PCR RFLP

目的建立检测高GC含量的CST3基因多态性的巢式PCR-RFLP方法.方法以巢氏PCR方法扩增CST3基因5′端和外显子1的特异性片段,用RFLP方法分析该基因的多态性.结果应用Enhancer System,以巢氏PCR方法能高效扩增出高GC含量的CST3基因5′端和外显子1区的特异性片段,用SacⅡ限制性酶切对CST3基因多态性进行有效分析.结论该方法能准确、高效的对CST3基因多态性进行分析,研究结果对高GC含量基因的研究有借鉴价值.

Objective To analyze CST3 gene polymorphism by nested PCR-RFLP, which with GC-rich （80% GC content）. Method The specificity fragment of 5＇ flanking region and exon lof CST3 was amplified with PCRX Enhancer System by Nested- PCR and analyzed by RFLP. Results The specificity fragment of CST3 with 80% GC content was amplified high efficiency and then analyzed CST3 gene polymorphism by restriction enzyme digestion with Sac Ⅱ. Conclusions The amplification efficiency of CST3 with GC-rich is improved greatly by Nested-PCR and with PCRX Enhancer System. CST3 gene polymorphism can be accurately analyzed by RFLP. The result can be used to research on genomics of GC-Rich.