花鲈微卫星标记分离及其多态性分析

Isolation of microsatellite markers for Lateolabrax japonicus and polymorphic analysis

该文利用FIASCO法(fast isolation by AFLP of sequences containing repeats)和GenBank数据库搜索法开发花鲈微卫星标记,并对筛选的标记进行多态性检测。两种方法共获得54条能够设计引物的序列,扩增结果显示15对引物具有多态性,多态性微卫星位点的等位基因数为2～10个。15个多态性位点中,4个位点偏离了Hardy-Weinberg平衡;各位点间没有连锁不平衡现象;仅位点SP52可能存在无效等位基因;除SP17和SP468外,其余引物的PIC值均在0.5以上,可用于花鲈群体遗传分析等研究。

To investigate population structure and marker assisted breeding,fast isolation by AFLP of sequences containing repeats（FIASCO） and GenBank database mining were used to develop novel microsatellite markers for sea perch（Lateolabrax japonicus）.Genomic DNA fragments containing SSR sequences were captured by hybridization to（GT）13 biotin-labeled probe and were ligated to PMD18-T vector.Among 150 randomly chosen clones from the SSR-enriched library,66 sequences contained microsatellite motif over five repeats.In addition,540 cDNA sequences and 132 ESTs of Lateolabrax japonicus were downloaded from GenBank and screened for di-,tri-and tetra-nucleotide repeats,while 22 sequences were found to contain microsatellites.As a result,15 microsatellite loci were shown to be polymorphic in 30 Lateolabrax japonicus individuals,with the alleles ranging from two to ten,the observed heterozygosities from 0.6000 ？ 1.0000,and the expected heterozygosities from 0.5079 ？ 0.8890.Four loci（SP17,SP52,SP94 and SP468） were deviated from HWE in the sampled population after Bonferroni＇s correction,and no linkage disequilibrium was found among all loci（P0.003）,whereas null alleles were detected at locus SP52（P0.05）.Among 15 polymorphic loci,the PIC values,which can be used for related population genetics analysis,were all above 0.5,with the exception of SP17 and SP468.