Villesen Palle, Fredsted Tina
Bioinformatics Research Center (BiRC), University of Aarhus, H, Guldbergsgade 10, building 1090, DK-8000 Aarhus C, Denmark.
BMC Ecol. 2006 Jun 8;6:8. doi: 10.1186/1472-6785-6-8.
One of the key tools for determining the social structure of wild and endangered primates is the ability to sex DNA from small amounts of non-invasive samples that are likely to include highly degraded DNA. Traditional markers for molecular sex determination of primates are developed on the basis of the human sequence and are often non-functional in distantly related primate species. Hence, it is highly desirable to develop markers that simultaneously detect Y- and X-chromosome specific sequences and also work across many species.
A novel method for sex identification in primates is described using a triple primer PCR reaction and agarose gel electrophoresis of the sex-chromosomal isoforms of the ubiquitously transcribed tetratricopeptide repeat protein gene (UTX/UTY). By comparing genomic data from several mammals we identified the UTX/UTY locus as the best candidate for a universal primate sexing marker. Using data from several species we identified a XY-conserved region, a Y conserved region and an X conserved region. This enabled the design of a triple primer PCR setup that amplifies X and Y products of different length in a single PCR reaction.
This simple PCR amplification of X and Y fragments is useful for sexing DNA samples from all species of primates. Furthermore, since the amplified fragments are very short the method can be applied to fragmented DNA extracted from non-invasive samples.
确定野生和濒危灵长类动物社会结构的关键工具之一,是能够从可能包含高度降解DNA的少量非侵入性样本中对DNA进行性别鉴定。传统的灵长类动物分子性别鉴定标记是基于人类序列开发的,在亲缘关系较远的灵长类物种中往往不起作用。因此,非常需要开发能够同时检测Y染色体和X染色体特异性序列且适用于多种物种的标记。
描述了一种利用三引物PCR反应和泛转录四肽重复蛋白基因(UTX/UTY)性染色体异构体的琼脂糖凝胶电泳进行灵长类动物性别鉴定的新方法。通过比较几种哺乳动物的基因组数据,我们确定UTX/UTY基因座是通用灵长类性别鉴定标记的最佳候选者。利用来自多个物种的数据,我们确定了一个XY保守区域、一个Y保守区域和一个X保守区域。这使得能够设计一种三引物PCR设置,在单个PCR反应中扩增不同长度的X和Y产物。
这种对X和Y片段进行简单的PCR扩增,可用于对所有灵长类物种DNA样本进行性别鉴定。此外,由于扩增片段非常短,该方法可应用于从非侵入性样本中提取的片段化DNA。