建立6种近交系大鼠遗传质量检测及品系鉴别的单核苷酸多态性组合

Establishment of Genetic Quality Testing and Strain Identification SNP panel for Six Classes Inbred Rats

目的 筛选出均匀分布在大鼠染色体上的单核苷酸多态性(SNP)位点,建立常用近交系大鼠遗传质量检测及品系鉴别方法。方法 在文献中获得SNP共141个,利用Ensembl数据库中获取的信息筛选出在BN、F344、GK、LEW、SHR和WKY 6种大鼠品系中呈现多态性的SNP位点,并从中优化出均匀分布各染色体且包含品系特异性SNP的最佳位点组合。通过PCR扩增技术和Sanger测序方法对组合位点用于常用近交系大鼠遗传质量检测和品系鉴别的效果进行验证。结果 优化出了在各品系内表现单态性、品系间呈多态性的40个SNP标记。其中包含可以用于品系鉴别的特异性位点:4个BN大鼠特异性位点,3个F344大鼠特异性位点,2个GK大鼠特异性位点,2个LEW大鼠特异性位点和1个SHR大鼠特异性位点。分别利用BN、F344、LEW、SHR和WKY大鼠的DNA混合样本对优化出的40个位点进行验证,结果与网站数据一致。结论 成功建立了常用近交系大鼠遗传质量检测及品系鉴别的SNP组合。

Objective To screen out single nucleotide polymorphism(SNP)loci evenly distributed on the rat chromosome that can be used to establish the genetic quality detection and strains identification of common inbred rats.Method A total of 141 SNPs were obtained from the literatures,and the information obtained in the Ensembl database was used to screen out SNPs with good polymorphism in 6 rat strains of BN,F344,GK,LEW,SHR and WKY.From the above SNPs,the SNP combination that evenly distributes each chromosome and contains strain-specific information was optimized.The effect of combined SNPs in genetic quality detection and strain identification of common inbred rats was verified by PCR amplification technology and Sanger sequencing method.Result 40 SNP markers were optimized,which were monomorphic within each strain and polymorphic among strains.They contain specific SNPs that can be used for strain identification,including 4 SNPs for BN rats,3 SNPs for F344 rats,2 SNPs for GK rats,2 SNPs for LEW rats,and 1 SNP for SHR rat.The optimized 40 SNPs were verified by using DNA mixed samples of BN rats,F344 rats,LEW rats,SHR rats and WKY rats respectively,and the result were consistent with the website data.Conclusion A SNP panel for genetic quality detection and strain identification of common inbred rats was successfully established.