【摘要】 采用FIASCO(Fast isolation 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。更多还原

【Abstract】 FIASCO(Fast isolation by AFLP sequences containing repeats)was used to screen microsatellite marker of Monopterus albus and analyze genetic variation of Monopterus albus populations with different genders.After enzyme cut,microsatellite probe hybridization and T vector linking,genome DNA of Monopterus albus was transformeded.into E.coli DH5a to construct Monopterus albus microsatellite-enriched libraries.Throuah sequencing of 75 positive colonies of random selection, 20 clones contained microsatellites were observed,and 15 pairs of primers were designed by Primer Premier V5.0.Through PCR of Monopterus albus genome DNA and agarose gel electrophoresis,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 malepopulation 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 results 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.更多还原