黄鳝微卫星标记筛选及其在不同性别表型群体中的遗传多态性  

Screening of Microsatellite and Genetic Polymorphism of Monopterus albus Populations with Different Gender Phenotype

作　　者：刘臻[1] 罗小华[1] 鲁双庆[1] 匡刚桥[1] 张建社[1] 

机构地区：[1]长沙大学生物工程与环境科学系,湖南长沙410003

出　　处：《江苏农业学报》2009年第2期333-338,共6页Jiangsu Journal of Agricultural Sciences

基　　金：国家自然科学基金项目(30571414);湖南省自然科学基金项目(06JJ20056);湖南省教育厅重点项目(08A007)

摘　　要：采用FIASCO（Fastisolation by AFLP sequences containing repeats）法进行黄鳝微卫星标记筛选并将其应用于黄鳝性别差异群体的遗传变异分析。黄鳝基因组DNA经酶切、微卫星核心序列探针杂交、T载体连接并转化至DH5a中，构建黄鳝基因组微卫星富集文库。随机挑选75个阳性克隆测序，结果发现其中20个含有微卫星序列，利用软件成功设计了15对微卫星引物，经过黄鳝基因组DNA的PCR扩增及电泳检测，筛选8对多态性微卫星引物并对黄鳝3种不同性别表型群体的基因组DNA进行PCR扫描，结果显示：在8个微卫星座位中，共检测到51个等位基因，平均每个座位的等位基因数为6．375个，等位基因频率分布为0．0012～0．6033；黄鳝雌性表型群体、间性表型群体、雄性表型群体的平均遗传杂合度分别为0．6371、0．6969、0．7345；平均多态信息含量分别为0．5656、0．6416、0．6858，表明这些微卫星标记在黄鳝不同性别表型群体的遗传多态性发生了改变，它们可作为黄鳝遗传背景分析的分子标记。FIASCO（ Fast isolation by AFLP sequences containing repeats） was used to screen mierosatellite marker of Monopterus albus and analyze genetic variation of Monopterus albus populations with different genders. After enzyme cut, mierosatellite probe hybridization and T vector linking, genome DNA of Monopterus albus was transformeded, into E. colt DHSa to construct Monopterus albus microsatellite-enriched libraries. Throuah sequencing of 75 positive colonies of random selection, 20 clones contained microsalellites were observed, and 15 pairs of primers were designed by Primer Premier VS. 0. Through PCR of Monopterus albus genome DNA and agarose gel eleetrophoresis, eight polymorphic microsatellite primers were screened out and PCR scan was carried out on genome DNA of three Monopterus albus populations with different genders. 51 alleles were found at 8 microsatellite loci,with average of 6. 375 alleles for each locus, and the frequency ranging from 0. 001 2 to 0. 603 3. The average heterozygosity of the female population, the neuter population and the male population were 0. 637 1, 0. 696 9, 0. 734 5,respectively, and the polymorphism information contents were 0. 565 6, 0. 641 6,0. 685 8, respectively. The resuhs indicated that the genetic polymorphism of these microsatellite markers of gender-based groups had changed, which could be used as molecular markers in the analysis of genetic background of Monopterus albus.

关 键 词：FIASCO 微卫星DNA 遗传多态性 

分 类 号：Q959.482[生物学—动物学]