滇西亚种树鼩微卫星分子标记的筛选  

Isolation and chatracterization of microsatellite markers in Tupaia belangeri chinensis

作　　者：张媛[1] 李晓飞[1] 李振宇[1] 仝品芬[1] 陈玲霞[1] 尹博文[1] 代解杰[1] 

机构地区：[1]中国医学科学院/北京协和医学院医学生物学研究所树鼩种质资源中心,昆明650118

出　　处：《中国比较医学杂志》2015年第6期36-41,共6页Chinese Journal of Comparative Medicine

基　　金：国家科技支撑计划项目(2014BAI01B01);云南省科技创新平台建设(2013DA002)

摘　　要：目的筛选出适用于滇西亚种树鼩种群遗传质量控制的特异性微卫星分子标记。方法首先从树鼩全基因组序列中筛选出约700个微卫星位点,择优选出约100个位点设计引物,去除有不良因素的,最后保留33对和文献报道的13对引物对滇西亚种树鼩DNA进行PCR扩增,根据琼脂糖电泳和聚丙烯酰胺电泳结果筛选保留,进行STR扫描再次筛选适于树鼩遗传检测的微卫星位点组合。结果筛选出树鼩微卫星位点22个,STR基因扫描有明显Stutter峰。比较备选位点和最终筛选出的位点,普通树鼩位点中选取5个,2个可用,与滇西亚种的重合率是40%;印度小树鼩位点中选取5个,2个可用,重合率是40%;中缅树鼩的位点中选取3个,2个可用,重合率约70%。结论筛选出的22个微卫星位点适用于滇西亚种树鼩的遗传检测,为树鼩种群的遗传质量监测提供科学依据。Objective To screen out specific microsatellite markers for use in Tupaia belangeri chinensis genetic testing. Methods Firstly to screen about 700 microsatellite loci from whole genome. Secondly to choose about 100 better loci without defect factors. Lastly 46 primers were designed by 33 tree shrew’s microsatellite loci obtained from whole genome and other references. Agarose gel electrophoresis and polyacrylamide gel electrophoresis were used for PCR products,and better loci based on electrophoresis results were chosen. Then STR scan was used to select the microsatellite loci combination for genetic testing. Results Twenty-two microsatellite loci were selected with a significant Stutter peak on STR scanning. Comparing the alternative loci and ultimately selected loci,there were two loci available in the five alternative loci of T. glis. The coincidence rate between T. glis and T. b. chinensis was 40%. There were two loci available in the five alternative loci of T. minor,and the coincidence rate between T. minor and T. b. chinensis was 40%. There were two loci available in the three alternative loci of T. belangeri,and the coincidence rate between T. belangeri and T. b.chinensis was about 70%. Conclusions The 22 microsatellite loci screened in this study are well applied for genetic testing of Tupaia belangeri chinensis,therefore,provide a scientific basis for the genetic quality monitoring of tree shrews.

关 键 词：滇西亚种树鼩 微卫星分子标记 STR扫描 

分 类 号：R[医药卫生]