双向等位基因特异性PCR技术在杏SNP分型上的应用  

Efficient procedure of Bi-directional PCR amplification of specific alleles for genotyping single nucleotide polymorphisms in apricot

作　　者：魏潇[1] 章秋平[1] 刘威生[1] 刘宁[1] 刘硕[1] 杨巍[1] 

机构地区：[1]辽宁省果树科学研究所.国家果树种质熊岳李杏圃,辽宁熊岳115009

出　　处：《果树学报》2013年第6期933-937,共5页Journal of Fruit Science

基　　金：国家自然科学基金项目(31071767);公益性行业(农业)科研专项(201003058)

摘　　要：【目的】建立一种基于单碱基突变的双向等位基因特异PCR(bi-directional PCR amplification of specific alleles,Bi-PASA)技术,并将其应用于杏单核苷酸多态性分型研究。【方法】利用直接测序的方法在2份不同果肉质地杏EXP基因的DNA序列上发现1个单碱基变异。在其上、下游两端设计双向等位基因特异性引物及其外侧互补引物,对120份杏材料的SNP进行分型。同时,对部分材料的基因型进行测序验证。【结果】利用双向等位基因特异PCR对杏EXP基因315 bp处SNP分型结果与直接测序完全一致。【结论】双向等位基因特异PCR技术是一种简单、经济、快速而可靠的SNP分型方法,可有效地应用于杏基因组上已知SNP突变的分型研究。[Objective]The objective of the study were to establish an bidirectional PCR amplification of specific alleles （BiPASA） technique system and to apply this technique to study single nucleotide poly morphism （SNP） of genes in apricot. [Method]A single nucleotide polymorphism was discovered by align ing the EXP genes of different texture eultivars in apricot. To type this SNP of 120 apricot cultivars, two outer （F and R） and two inner allelespecific （P and Q） primers were designed using the SNP as the 3＂ end. In addition, we studied the assay feasibility of BiPASA to SNPs identification by sequencing. [Result] The results from the BiPASA assay were completely consistent with those of sequencing. [Conclusion] BiPASA is a simple, economical, rapid and reliable new method for SNP typing particularly for detecting known mutations in apricot.

关 键 词：杏 单核苷酸多态性 双向等位基因特异PCR 

分 类 号：S662.2[农业科学—果树学]