豚鼠个体及品系间多态性微卫星位点筛选
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浙江省公益技术应用研究(实验动物)计划项目(编号:2017C37144);卫生部科学研究基金(编号:98-2-323)。


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

    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。

    Abstract:

    Objective To screen the polymorphism microsatellite markers of guinea pigs, and provide basic information for identification of genetic structure and gene mapping in guinea pigs. Methods The genomic DNA of guinea pigs was found out on foreign websites, and microsatellite sequences characterized by AC/GT were selected. A total of 400 pairs 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. Results At the first step, 10 DNA samples from 5 strains of guinea pigs were taken for PCR, and 110 pairs of polymorphic microsatellite markers with 1-3 electrophoretic bands were screened from the designed 400 pairs of primers. At the second step, 60 DNA samples from 5 strains of guinea pigs were assayed by PCR and 51 pairs of polymorphic primers with high resolution electrophoresis bands were screened from the above 110 pairs of primers. Among these screened 51 primers, more than 10 pairs of primers showed accordant or mostly accordant differences among strains. Conclusions The 51 pairs 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.

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卫振,洪胜辉,刘迪文.豚鼠个体及品系间多态性微卫星位点筛选[J].中国比较医学杂志,2017,27(11):32~37.

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  • 收稿日期:2017-03-17
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  • 在线发布日期: 2017-11-28
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