一种基于变性高效液相色谱技术筛选单核苷酸多态性并估算基因频率的方法

A method for estimating allelic frequencies of single nucleotide polymorphisms (SNPs) based on denaturing high performance liquid chromatography (DHPLC)

选取生产性能具有明显差异的4个鸡品种(莱航鸡、隐性白洛克鸡、丝羽乌骨鸡和杏花鸡)为材料,利用变性高效液相色谱(DHPLC)技术检测鸡生长激素基因部分序列(518bp)的多态性,快速筛查到5个可能与生产性能相关的单核苷酸多态性(SNPs).根据DHPLC杂合谱型与测序基因型一一对应的关系,介绍了当所检测目的DNA片段中含有多个SNPs时基因频率估算的简单方法,比较估算的和PCR RFLP统计的基因频率,发现差异不显著,证明该估算方法有一定的可行性.

Four breeds of chickens (white Leghorn, white recessive rocks, Taihe silkies and Xinhua) with different production performance were applied to screen potential single nucleotide polymorphisms (SNPs) related to production traits in a region of the growth hormone gene using denaturing high performance liquid chromatography (DHPLC). Five SNPs were found in a fragment of 518 bp. In the present study a simple and rapid method to estimate allelic frequencies of SNPs in the case of several SNPs existing in the same targeted DNA fragment was introduced based on the correspondence between the heterozygous DHPLC peak and sequencing genotype, of which the accuracy was evaluated by comparing with the allelic frequencies result of PCR-RFLP. It's a feasible method because no significant difference was detected between the estimated allelic frequencies and those calculated by PCR-RFLP.