等位基因特异性引物PCR技术及其应用研究

Study of Allelic Specific Primer Polymerase Chain Reaction Technique and its Application

目的 :研究建立等位基因特异性引物PCR技术体系 ,并将其应用于基因单核苷酸多态性研究工作。方法 :通过美国国家生物信息中心 (NCBI)的genBank获取基因序列及其相应位点的SNP信息。利用Primer5 .0软件设计引物 ,并经NCBI的Blast2 .0软件检验其特异性。结果 :建立了单一等位基因特异性引物PCR(SASP -PCR)与嵌套式等位基因特异性引物PCR(NASP—PCR)两种技术 ,并应用于 β2 肾上腺素受体及内皮源性一氧化氮合酶基因单核苷酸多态性的研究 ,证实该技术的稳定性和优越性。结论 :等位基因特异性引物PCR技术是一种更为简便、特异性较高、费用少的、便于推广的SNP检测方法 。

Objective:To study and establish an allelic specific primer polymerase chain reaction (ASP-PCR)technique system and to apply this technique to study single nucleotide polymorphism(SNP) of genes.Methods:It got the information of nucleotide sequences and their corresponding SNPs of gene from genBank of National Central of Biotechnology Information(NCBI).The primers were designed by Primer 5.0 software and their specificity was tested by NCBI Blast 2.0 software.Results:They successfully designed and established two technique systems of single allelic specific primer polymerase chain reaction (SASP-PCR)and nested allelic specific primer polymerase chain reaction(NASP-PCR.The two methods were applied to study SNPs of beta-2 Adrenoceptor and endothelium nitric oxide synthase.The superiority and stability of ASP-PCR technique on determining SNPs of genes have been tested.Conclusions:The ASP-PCR technique is a new method on determination of SNP with more convenient,more specific,less spend and easy to application and has much more advantage especially in the study of population gene polymorphism.