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| Characterization Analysis of SLA-1 Gene from SPF Yorkshire (Sus scrofa) and Landrace |
| 1, 1, 1, 1, 1, 2, 1 |
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2. Gansu Agricultural University
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Abstract Abstract Swine (Sus scrofa) leukocyte antigen (SLA) gene is one of the genetic factors that cause xenotransplantation cell immune rejection. To enrich the classical experimental animal model and solve the problem of the lack of heterotopic transplantation model, blood samples were obtained from specific pathogen free (SPF) Yorkshire and Landrace for isolation of peripheral blood mononuclear cells, and SLA-1 gene was amplified by PCR after reverse transcription. SLA-1 gene sequences were obtained by analyzing the direct and cloning results. The polymorphism of alleles was analyzed using the bioinformatics software. A total of 16 alleles was identified at SLA-1 gene, containing a full-length 1 086 bp open reading frame. Nucleotide diversity (Pi) was 0.045 59, haplotype diversity (Hd) was 0.000 49, the average nucleotide difference (K) was 49.508, and the content of G+C reached 60.9%. Among 150 nucleotide mutations, the number of nucleotide parsimony informative sites (NPi) were higher than those of the nucleotide singleton variable sites (NS), and 82 of the 361 amino acids were mutated and the numbers of non-synonymous sites (NSS) were obviously higher than those of synonymous sites (SS). The polymorphism of SLA-1 allele was mainly concentrated in exon 2 and 3. Nine of 33 key amino acids were completely conserved, and eight residues showed complete consistence with the corresponding amino acids of human leukocyte antigen (HLA) allele. Eleven of the 19 amino acids, which bind β2-microglobulin (β2m) to HLA-A, showed complete consistency among the alleles identified in this study. The key amino acid binding sites of CD8 to major histocompatibility complex (MHC) were highly consistent in human (Homo sapiens) and SPF Yorkshire and Landrace, except for 225 (Thr/Ser) and 228 (Thr/Met) sites. The result of phylogenetic relationship showed that there was a close relationship among SPF Yorkshire and Landrace pigs, Korean native pigs and Meishan pigs. This study can provide genetic basis for establishment of animal models antigen presentation and immune response.
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Received: 12 December 2016
Published: 02 May 2017
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