用LA-PCR方法克隆东方田鼠部分Y染色体序列  

Exploring SNPs on ChrY of Microtus fortis

作　　者：姜宪环[1] 蔡慧强[1] 高骏[2] 倪丽菊 李凯[1] 肖君华[1] 周宇荀[1] 

机构地区：[1]东华大学生物科学与技术研究所,上海201620 [2]上海市农业科学院,上海201106 [3]上海实验动物研究中心,上海201203

出　　处：《实验动物与比较医学》2012年第4期324-328,333,共6页Laboratory Animal and Comparative Medicine

基　　金：上海市科技创新行动计划实验动物专项基金

摘　　要：目的克隆东方田鼠长江亚种（Microtus fortis calamorum）和宁夏指名亚种（Microtus fortis fortis）Y染色体的部分序列，并挖掘这两个亚种间的单核苷酸多态性位点（SNPs）。方法本研究利用依据Y染色体保守性靶标位点（YCATS）设计的引物，测得Y染色体上的8个内含子短片段序列，进而设计长片段PCR（LA—PCR）引物来获得相距几kb的两个内含子之间的长片段序列。结果获得东方田鼠长江亚种和宁夏指名亚种Y染色体上的7300bp序列，比对这两个亚种的序列后发现4个SNPs位点。结论获得了田鼠Y染色体上的7300bp序列和4个SNPs位点。Objective To obtain the sequences of Y chromosome of Microtus fortis calamorum and Microtusfortisfortis, and find out some SNPs of these two subspecies. Methods Eight short segments＇ sequences involving in introns of ChrY are sequenced by using Y chromosome conserved anchored tagged primers, which would be used to design two new pairs of primers to achieve two long fragments of several kb in size through LA-PCR （Long and accurate polymerase chain reaction）. Results About 7300 bp long on ChrY of M.f.calamorum and M.ffortis is obtained respectively, for the first time, and 4 SNPs by comparing the sequences of these two subspecies are obtained. Conclusion The 7300 bp sequences on Y chr and 4 SNPs are obtained.

关 键 词：东方田鼠 单核苷酸多态性位点 Y染色体保守性靶标位点 长片段PCR 

分 类 号：Q95-33[生物学—动物学]