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MBE Advance Access published online on December 15, 2004
Molecular Biology and Evolution, doi:10.1093/molbev/msi053
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Molecular Biology and Evolution © Society for Molecular Biology and Evolution 2004; all rights reserved.
Accepted October 26, 2004

Research Article

A Universal Primer Set for PCR amplification of Nuclear Histone H4 Genes from all Animal Species

Pascal Pineau 1*, Michel Henry 2, Rodolphe Suspène 2, Agnès Marchio 1, Agnès Dettai 3, Régis Debruyne 4, Thierry Petit 5, Alexis Lécu 6, Pierre Moisson 7, Anne Dejean 1, Simon Wain-Hobson 2, and Jean-Pierre Vartanian 2

1 Unité d'Organisation Nucléaire et Oncogénèse, INSERM U579, Institut Pasteur
2 Unité de Rétrovirologie Moléculaire URA 1930, Institut Pasteur
3 Service de Systématique Moléculaire, Institut de Systématique, Museum National d'Histoire Naturelle, Paris
4 UMR 5143 CNRS, Paléobiodiversité, Museum National d'Histoire Naturelle, Paris
5 Zoo de la Palmyre, Les Mathes
6 Services Vétérinaires, Zoo de Vincennes, Museum National d'Histoire Naturelle, Paris
7 Jardin Zoologique, Mulhouse, France

* To whom correspondence should be addressed.
Pascal Pineau, E-mail: ppineau{at}pasteur.fr


  Abstract

In order to control the quality of genomic DNA of samples from a wide variety of animals, a hemi-nested 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, 8 samples from invertebrates and 35 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, 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.

Keywords: histone H4; PCR; vertebrates.
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