鉴定清道夫受体BI基因敲除突变小鼠的新方法

A novel method for genotyping scavenger receptor BI gene knockout mice

目的建立一种清道夫受体BI(SR-BI)基因敲除突变小鼠基因型的鉴定方法。方法参照碱基淬灭探针技术原理,设计鉴定野生型和突变型SR-BI基因的引物探针。反应体系经PCR扩增后,利用融解曲线功能判断小鼠基因型。结果新方法可在1h内完成基因型分析。野生型和突变型纯合子小鼠分别在(53.25±0.81)℃及(62.25±0.31)℃处出现单个融解谷;而杂合子小鼠在两处均出现融解谷。结论新方法简单、快速、准确,适用于鉴别SR-BI基因敲除突变小鼠的基因型。

Objective To establish a practical method for determining the genotype of scavenger receptor BI（SR-BI）gene knockout mice.Methods Primers and probes were designed according to the base-quenched probe technique and the targeted SR-BI genotypes were analyzed using melting curve.Results The analysis for genotyping could be completed with the tecnigue mentioned above within 1 h.Single melting valley appeared at the melting temperatures of （62.25±0.31）℃ and （53.25±0.81）℃ for the rats with homozygous and wild-type,respectively.The rats with heterozygous showed two melting valleys.Conclusion The procedure of this novel method is accurate,simple and quicke,which is suitable for genotyping the SR-BI gene knockout mice.