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东方田鼠微卫星标记的富集筛选与初步应用
引用本文:倪丽菊,陶凌云,柏熊,胡建华,高诚,谢建云.东方田鼠微卫星标记的富集筛选与初步应用[J].遗传,2011,33(9):989-995.
作者姓名:倪丽菊  陶凌云  柏熊  胡建华  高诚  谢建云
作者单位:1. 上海实验动物研究中心,上海 201203;上海市实验动物质量监督检验站,上海 201203
2. 上海西普尔-必凯实验动物有限公司,上海,201203
基金项目:上海市科委基金项目(编号:071409001,09140900101)资助
摘    要:根据生物素与链霉亲和素的亲和原理,利用磁珠富集法筛选东方田鼠(Microtus fortis)微卫星分子标记.链霉亲和素磁珠捕获生物素标记的微卫星探针,然后与连有接头的单链限制性酶切片段复性结合,获得含有微卫星的单链片段,PCR扩增形成双链,连接T载体并转化感受态细胞,得到东方田鼠微卫星富集文库.随机挑选70个阳性克隆,经测序分析,获得微卫星序列92个.设计合成27对微卫星引物并成功筛选出21对可用引物,取其中10对引物,荧光标记后对3个人工驯养及野生东方田鼠种群进行遗传多样性分析.结果显示,文章所构建的东方田鼠微卫星文库的阳性克隆率较高,初步筛选的10个微卫星标记均为具有高度多态性的微卫星标记.在3个东方田鼠种群中,野生湖南种群的观测等位基因数(Na)、有效等位基因数(Ne)、观测杂合度(Ho)、期望杂合度(He)和多态信息含量(PIC)均最高,人工驯养的湖南种群次之,人工驯养的宁夏种群最低.

关 键 词:东方田鼠  磁珠富集  微卫星标记  遗传多样性

Enrichment and screening of the microsatellite markers in Microtus fortis and its preliminary application in genetic diversity research
NI Li-Ju,TAO Ling-Yun,BAI Xiong,HU Jian-Hua,GAO Cheng,XIE Jian-Yun.Enrichment and screening of the microsatellite markers in Microtus fortis and its preliminary application in genetic diversity research[J].Hereditas,2011,33(9):989-995.
Authors:NI Li-Ju  TAO Ling-Yun  BAI Xiong  HU Jian-Hua  GAO Cheng  XIE Jian-Yun
Institution:NI Li-Ju1,2,TAO Ling-Yun1,BAI Xiong3,HU Jian-Hua1,GAO Cheng1,XIE Jian-Yun1,2 1.Shanghai Laboratory Animal Research Center,Shanghai 201203,China,2.Shanghai Quality Monitoring Center for Laboratory Animal,3.Shanghai SIPPR-BK LAB Animal ltd
Abstract:This study was to isolate microsatellite markers from Microtus fortis genome by magnetic beads enrichments.Through hybridization of biotin-labeled microsatellite oligonucleotide probes,which were captured by streptavidin-coated magnetic with the adaptor-ligated enzyme-digested genome fragments,single-stranded DNA fragments containing microsatellites were obtained.After PCR amplification,these fragments were then cloned into T vectors and were transformed into competent cells subsequently.Ninety-two microsat...
Keywords:Microtus fortis  enrichment by magnetic beads  microsatellite  genetic diversity  
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