Hare M P, Karl S A, Avise J C
Department of Genetics, University of Georgia, Athens 30602, USA.
Mol Biol Evol. 1996 Feb;13(2):334-45. doi: 10.1093/oxfordjournals.molbev.a025593.
A puzzling population-genetic phenomenon widely reported in allozyme surveys of marine bivalves is the occurrence of heterozygote deficits relative to Hardy-Weinberg expectations. Possible explanations for this pattern are categorized with respect to whether the effects should be confined to protein-level assays or are genomically pervasive and expected to be registered in both protein- and DNA-level assays. Anonymous nuclear DNA markers from the American oyster were employed to reexamine the phenomenon. In assays based on the polymerase chain reaction (PCR), two DNA-level processes were encountered that can lead to artifactual genotypic scorings: (a) differential amplification of alleles at a target locus and (b) amplification from multiple paralogous loci. We describe symptoms of these complications and prescribe methods that should generally help to ameliorate them. When artifactual scorings at two anonymous DNA loci in the American oyster were corrected, Hardy-Weinberg deviations registered in preliminary population assays decreased to nonsignificant values. Implications of these findings for the heterozygote-deficit phenomenon in marine bivalves, and for the general development and use of PCR-based assays, are discussed.
在海洋双壳贝类的等位酶调查中广泛报道的一个令人费解的群体遗传现象是,相对于哈迪-温伯格预期,杂合子出现不足。根据这些影响是应局限于蛋白质水平检测还是在基因组中普遍存在并预期在蛋白质和DNA水平检测中都能体现,对这种模式的可能解释进行了分类。利用美国牡蛎的匿名核DNA标记重新审视了这一现象。在基于聚合酶链反应(PCR)的检测中,遇到了两种可能导致人为基因型评分的DNA水平过程:(a)目标位点上等位基因的差异扩增,以及(b)来自多个旁系同源位点的扩增。我们描述了这些并发症的症状,并给出了通常有助于缓解它们的方法。当美国牡蛎两个匿名DNA位点的人为评分得到纠正后,初步群体检测中记录的哈迪-温伯格偏差降至不显著的值。讨论了这些发现对海洋双壳贝类杂合子不足现象以及基于PCR检测的一般开发和使用的影响。