荷斯坦奶牛脊椎畸形综合征焦磷酸测序检测方法建立与应用

摘要

为建立一种快速高通量的荷斯坦奶牛脊柱畸形综合征（complex vertebral malformation，CVM）的分子检测方法，根据CVM是由于SLC35A3基因第4外显子559位处发生G→T突变的遗传学基础，利用Assay Design SW软件设计了1对PCR引物和1条测序引物对三种不同基因型的基因材料进行测序分析，建立焦磷酸测序检测方法。对55份进境荷斯坦奶牛血样进行检测，发现有1例为隐性基因携带者，将PCR产物进行测序，其DNA序列与焦磷酸测序检测结果一致。研究表明该方法是一种有效的新方法，可用于荷斯坦奶牛脊椎畸形综合征的检测。%To develop a rapid and high-flux molecular method for detection of complex vertebral malformation (CVM),one pair of PCR primers and one sequencing primer were designed according to the gene sequence of bovine SLC35A3 gene by the Assay Design SW software. The PCR products of three different genotypes were detected via pyrosequencing method. There was one CVM carrier detected in 55 blood samples from imported Holstein bovine population. The pyrosequencing result was confirmed by sequencing of PCR product of the CVM carrier.