豚鼠个体及品系间多态性微卫星位点筛选

Screening of microsatellite loci with polymorphism among individuals and strains of guinea pigs

目的筛选豚鼠多态性微卫星标记,为鉴定豚鼠遗传结构及基因定位提供基础信息。方法从国外网站筛查得豚鼠基因组DNA资料,选择AC、GT为核心的微卫星序列,根据其旁侧序列用软件设计400对引物。经过二轮PCR及电泳等实验,以电泳条带作为遗传标记,通过豚鼠微卫星结构的评价筛选出多态性引物。结果第一步用5个品系豚鼠的10份DNA样本进行PCR,从400对引物中筛选出约110对具有多态性的豚鼠引物,其产物电泳条带数为1~3条。第二步用5个品系豚鼠的60份DNA样本进行PCR,从上述110对引物中筛选出51对电泳条带分辨率高的多态性引物,其中10余对引物在品系间呈标记一致或多数一致的差异。结论筛选出的51对引物在个体及品系间呈多态性,未见其他报道,可用于常规检测研究,部分可能用于豚鼠性状基因定位。

ObjectiveTo screen the polymorphism microsatellite markers of guinea pigs,and provide basic information for identification of genetic structure and gene mapping in guinea pigs.MethodsThe genomic DNA of guinea pigs was found out on foreign websites,and microsatellite sequences characterized by AC/GT were selected.A total of400pairs of primers were designed using software according to their flanking sequences.After two rounds of PCR and electrophoresis,with the electrophoresis bands serving as genetic markers,the polymorphic primers were screened by the evaluation of microsatellite structure of guinea pigs.ResultsAt the first step,10DNA samples from5strains of guinea pigs were taken for PCR,and110pairs of polymorphic microsatellite markers with1-3electrophoretic bands were screened from the designed400pairs of primers.At the second step,60DNA samples from5strains of guinea pigs were assayed by PCR and51pairs of polymorphic primers with high resolution electrophoresis bands were screened from the above110pairs of primers.Among these screened51primers,more than10pairs of primers showed accordant or mostly accordant differences among strains.ConclusionsThe51pairs of primers are polymorphic among individuals and strains,and have not been found in other reports so far.They can be used for conventional genetic detection and related studies,and some of them can be used for gene mapping of the traits in guinea pigs.