IUCN / SSC Cat Specialist Group - Digital Cat Library |
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| Wang, Q.; Wu, X.-B.; Yan, P.; Zheng, S. | |
| Sequence variability analysis on major histocompatibility complex class II DRB alleles in three felines | |
| 2008 Frontiers of Biology in China (3): 55-62 | |
The variation of the exon 2 of the major histocompatibility complex (MHC) class II gene DRB locus in three feline species were examined on clouded leopard (_Neofelis nebulosa_), leopard (_Panthera pardus_) and Amur tiger (_P. tigris altaica_). A pair of degenerated primers was used to amplify DRB locus covering almost the whole exon 2. Exon 2 encodes the รก1 domain which is the most variable fragments of the MHC class II molecule. Single-strand conformational polymorphism (SSCP) analysis was applied to detect different MHC class II DRB haplotypes. Fifteen recombinant plasmids for each individual were screened out, isolated, purified and sequenced finally. Totally eight distinct haplotypes of exon 2 were obtained in four individuals. Within 237 bp nucleotide sequences from four samples, 30 variable positions were found, and 21 putative peptide-binding positions were disclosed in 79 amino acid residues. The ratio of nonsynonymous substitutions (dN) was much higher than that of synonymous substitutions (dS), which indicated that balancing selection probably maintain the variation of exon 2. MEGA neighbor joining (NJ) and PAUP maximum parsimony (MP) methods were used to reconstruct phylogenetic trees among species, respectively. Results displayed a more close relationship between leopard and tiger; however, clouded leopard has a comparatively distant distant relationship from the other two. |
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