Pineau Pascal, Henry Michel, Suspène Rodolphe, Marchio Agnès, Dettai Agnès, Debruyne Régis, Petit Thierry, Lécu Alexis, Moisson Pierre, Dejean Anne, Wain-Hobson Simon, Vartanian Jean-Pierre
Unité d'Organisation Nucléaire et Oncogénèse, INSERM U579, Institut Pasteur, Paris, France.
Mol Biol Evol. 2005 Mar;22(3):582-8. doi: 10.1093/molbev/msi053. Epub 2004 Dec 15.
To control the quality of genomic DNA of samples from a wide variety of animals, a heminested PCR assay specifically targeting a nuclear gene has been developed. The histone H4 gene family comprises a small number of genes considered among the most conserved genes in living organisms. Tissue samples from necropsies and from cells belonging to 43 different species were studied, eight samples from invertebrates and 35 samples from vertebrates covering all classes. Ancient DNA samples from three Siberian woolly mammoths (Mammuthus primigenius) dating between 40,000 and 49,000 years before present were also tested for PCR amplification. Performance of HIST2H4 amplification were also compared with those of previously published universal PCRs (28S rRNA, 18S rRNA, and cytochrome b). Overall, 95% of species studied yielded an amplification product, including some old samples from gorilla and chimpanzees. The data indicate that the HIST2H4 amplimers are, thus, suitable for both DNA quality testing as well as species identification in the animal kingdom.
为了控制来自多种动物样本的基因组DNA质量，已开发出一种专门针对核基因的半巢式PCR检测方法。组蛋白H4基因家族由少数被认为是生物中最保守的基因组成。研究了来自尸检的组织样本以及属于43个不同物种的细胞样本，其中有8个无脊椎动物样本和35个涵盖所有类别的脊椎动物样本。还对来自距今40000至49000年前的三头西伯利亚猛犸象（猛犸象）的古代DNA样本进行了PCR扩增测试。还将HIST2H4扩增的性能与先前发表的通用PCR（28S rRNA、18S rRNA和细胞色素b）的性能进行了比较。总体而言，95%的研究物种产生了扩增产物，包括一些来自大猩猩和黑猩猩的旧样本。数据表明，因此，HIST2H4扩增子适用于动物王国中的DNA质量检测以及物种鉴定。
