微卫星DNA标记在近交系大鼠HFJ和MIJ遗传监测中的应用

Microsatellite DNA analysis for genetic monitoring of inbred strain HFJ and MIJ rats

目的用24对引物对近交系HFJ和MIJ大鼠的微卫星位点进行多态性分析,并选用近交系Lewis和F344大鼠作为对照,进行比较分析。方法用传统的酚-氯仿法分别提取4个近交系大鼠MIJ、HFJ、Lewis和F344的基因组DNA,选取大鼠24个微卫星位点,通过PCR扩增,扩增产物经过非变性聚丙烯酰胺凝胶电泳和银染,根据电泳结果,比较分析4种品系近交系大鼠之间微卫星多态性。结果 4种品系及品系内不同个体的近交系大鼠在24个微卫星位点上的扩增产物均出现一个条带,MIJ和HFJ大鼠在品系间和品系内均表现为单态性,同Lewis和F344的扩增结果比较,14个位点显示多态性,有10个位点显示单态性。结论两个近交系大鼠品系MIJ和HFJ符合近交系要求,筛选出的14个多态性微卫星位点可用于有关近交系大鼠的遗传背景监测。

Objective The polymorphism of microsatellite loci among four inbred strains of rats（HFJ,MIJ,Lewis,F344） was analyzed using 24 pair primers.Methods To extract the genomic DNA of MIJ,HFJ,Lewis and F344 inbred rats by standard extraction procedure with phenol/chloroform.24 microsatellite loci were selected for PCR amplification.Through native polyacrylamide gel electrophoresis and silver staining,microsatellite polymorphism was analyzed in the four strains of inbred rats.Results The amplified products of 24 microsatellite loci among different individual rats of these 4 strains and the same strain rats displayed single allelic gene band.Among the 24 microsatellite loci,it displayed monomorphism between HFJ and MIJ rats.But HFJ and MIJ rats had significant deviations from Lewis and F344 rats.14 microsatellite loci turned out to be polymorphic,and 10 microsatellite loci displayed monomorphism.Conclusions The results of this study suggest that MIJ and HFJ strains basically meet the requirement of inbred strain.The 14 screened loci can be used for genetic monitoring of these inbred rats.