Tang S, Tassanakajon A, Klinbunga S, Jarayabhand P, Menasveta P
Department of Biochemistry, Faculty of Science, Chulalongkorn University, Bangkok, 10330, Thailand.
Mar Biotechnol (NY). 2004 Nov-Dec;6(6):604-11. doi: 10.1007/s10126-004-2300-5.
Three partial genomic libraries were constructed from genomic DNA of the tropical abalone (Haliotis asinina) that was digested with AluI, vortexed/sonicated, and digested with mixed enzyme (AluI, HincII, and RsaI). The libraries yielded 0.02%, 0.42%, and 1.46% positive microsatellite-containing clones, respectively. Eleven clones each of perfect, imperfect, and compound microsatellites were isolated. Ten primer pairs (CUHas1-CUHas10) were analyzed to evaluate their polymorphic level. The numbers of alleles per locus, observed heterozygosity (H0), and expected heterozygosity (He) ranged from 3 to 26 alleles, and varied between 0.27 and 0.85 and between 0.24 and 0.93, respectively. Three microsatellite loci (CUHas2, CUHas3, and CUHas8) were further used for examination of genetic diversity and differentiation of natural H. asinina in coastal waters of Thailand. Genetic variabilities in terms of the effective number of alleles (n(e)), H0, and He were higher in 2 samples from the Gulf of Thailand (n(e)=9.37, 7.66; H0=0.62, 0.78; and He=0.87, 0.86) than those of one sample (n(e)=6.04; H0=0.58; and He=0.62) derived from the Andaman Sea. Assessment of genetic heterogeneity, including allele frequency comparison and pairwise F(ST) analysis, indicated interpopulational differentiation, between natural H. asinina from the Gulf of Thailand and that from the Andaman Sea (P<0.0001).
从热带鲍鱼(Haliotis asinina)的基因组DNA构建了三个部分基因组文库,该基因组DNA先用AluI酶切,涡旋/超声处理,再用混合酶(AluI、HincII和RsaI)酶切。这些文库分别产生了0.02%、0.42%和1.46%含微卫星的阳性克隆。分离出了11个完美型、非完美型和复合型微卫星克隆。分析了10对引物(CUHas1 - CUHas10)以评估其多态性水平。每个位点的等位基因数、观察杂合度(H0)和期望杂合度(He)分别为3至26个等位基因,H0在0.27至0.85之间变化,He在0.24至0.93之间变化。进一步使用三个微卫星位点(CUHas2、CUHas3和CUHas8)来检测泰国沿海水域天然H. asinina的遗传多样性和分化。泰国湾的两个样本在有效等位基因数(n(e))、H0和He方面的遗传变异性(n(e)=9.37, 7.66;H0=0.62, 0.78;He=0.87, 0.86)高于来自安达曼海的一个样本(n(e)=6.04;H0=0.58;He=0.62)。包括等位基因频率比较和成对F(ST)分析在内的遗传异质性评估表明,泰国湾的天然H. asinina与安达曼海的天然H. asinina之间存在种群间分化(P<0.0001)。