利用等位基因特异性PCR技术检测番茄高色素基因hp1和hp2的SNP

摘要：番茄高色素基因hp1和hp2突变体与野生型基因相比均发生了单碱基的点突变，常规PCR技术很难把它们区分开。本试验对碱基错配类型和位置与特异性PCR的关系进行了研究。结果表明：引入TA-TG双碱基错配的引物，退火温度在49.3+0.1℃时能把hp1基因的突变位点和野生型的对应位点区分开来。引入T-C碱基错配的引物，退火温度在53.5+0.1℃时能把hp2基因的突变位点和野生型的对应位点区分开来。错配碱基在引物3′端的位置对PCR的特异性影响不大。

Assaying Single Nucleotide Polymorphism of High Pigment Genes hp1 and hp2 Based on Allele-specific PCR in Tomato

Abstract: Comparing with their corresponding wild allele genes, a single nucleotide mutation had been taken place in high pigment genes hp1 and hp2, respectively, which was very difficult to be discriminated by normal PCR technique. In this experiment, the relationship between specific PCR and mismatched bases as well as their positions were studied. Results showed that the primer with double-base mismatch TA-TG can distinguish mutation location of hp1 from its wild allele location at annealing temperature 49.3 +0.1 ℃. A primer with T-C mismatch can distinguish mutation location of hp2 from its wild allele location at annealing temperature 53.5+0.1 ℃. The postion of mismatched bases at 3` end of primers had little effect on specificity of PCR.