Abstract
The MHC class II genes of many primate species were investigated extensively in recent years. However, while Mhc-DRB genes were studied in Old World monkeys such as rhesus macaques, the Mhc-DRB of baboons was only studied in a limited way. Because of their close anatomical and physiological relationship to humans, baboons are often used as models for reproduction and transplantation research. Baboons are also studied as a model species in behavioural ecology. Thus, identification of MHC genes would provide a foundation for studies of Mhc, biology and behaviour. Here, we describe the use of PCR, cloning, denaturing gradient gel electrophoresis (DGGE) and sequencing to identify Mhc-DRB sequences in wild chacma baboons (Papio ursinus). We amplified the highly variable second exon of baboon Mhc-DRB sequences using generic DRB primers. To validate and optimize the DGGE protocol, four DNA samples were initially studied using cloning and sequencing. Clones were screened using a novel RFLP approach to increase the number of clones identified for each individual. Results from cloning and sequencing were used to optimise DGGE conditions for Mhc-DRB genotyping of the remaining study subjects. Using these techniques, we identified 16 Paur-DRB sequences from 30 chacma baboons. On the basis of phylogenetic tree analyses, representatives of the Mhc-DRB1 and Mhc-DRB5 loci, and 13 different DRB lineages were identified. Evidence for trans-species inheritance of some Mhc-DRB sequences comes from high identity between the new Paur-DRB sequences and sequences from Papio cynocephalus, Macaca mulatta and possibly Galago moholi.
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Acknowledgements
For the assistance with sample collection in the field, we gratefully thank all those individuals who helped with our trapping operations in 2000 and 2001, especially Crispian Barlow, Andre and Tana Burger, Fredi Devas, Anna Randall, H.O. Reuter, Marcus Rowcliffe, and Hartmut Winterbach. Special thanks also go to Anna Randall and Stephan Funk for conducting the DNA extractions (with additional assistance from Dada Gotelli) in London. We also thank Polla and Chrija Swart for their permission to work at the Tsaobis Leopard Park, the Desert Research Foundation of Namibia for affiliation, and the Ministry of Environment and Tourism for research permission. We confirm that we have adhered to the Guidelines for the Use of Animals in Behavioural Research and Teaching (Animal Behaviour 2003, 65:249–255) and the legal requirements of the country in which the work was carried out (Namibia). This work was funded by a Natural Environment Research Council (UK) grant awarded to GC, and a Ministère de l’Education et de la Recherche (France) Studentship awarded to EH. GC was funded by a NERC Advanced Fellowship during the writing of this paper. This paper is a publication of the ZSL Institute of Zoology’s Tsaobis Baboon Project. Contribution ISEM 2006, #XXX.
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Nucleotide sequence data reported are available in the GenBank/EMBL/DDBJ databases under the accession numbers DQ339722–DQ339737.
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Huchard, E., Cowlishaw, G., Raymond, M. et al. Molecular study of Mhc-DRB in wild chacma baboons reveals high variability and evidence for trans-species inheritance. Immunogenetics 58, 805–816 (2006). https://doi.org/10.1007/s00251-006-0156-8
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DOI: https://doi.org/10.1007/s00251-006-0156-8