实验用荣昌猪SLA-1等位基因鉴定及分子遗传特征分析

doi:10.11843/j.issn.0366-6964.2023.07.037

Abstract

Abstract: The study aimed to clarify the SLA genetic background of experimental Rongchang pigs and further study SLA-1 molecule-related antigen presentation and immune response mechanism. In this study, the anticoagulant bloods were collected from 27 Rongchang pigs, and peripheral blood lymphocytes were isolated. Total RNA were extracted and SLA-1 genes were amplified using specific primers with RT-PCR method, cloned and sequenced. Molecular genetic characte-ristics of the obtained sequences were further analyzed. The results showed that a total of 11 SLA-1 alleles were obtained in Rongchang pigs, of which nine were novel alleles. All alleles obtained GenBank accession numbers and official names were assigned by the ISAG SLA Nomenclature Committee. The frequency of SLA-1^*24:01 allele was the highest in this population. The full-length of the coding region of SLA-1 gene was 1 086 bp, with 127 nucleotide polymorphic sites. The number of non-synonymous SNPs was higher than that of synonymous SNPs. The nucleotide diversity was 0.044 9, the haplotype diversity was 1, the average nucleotide difference number was 48.782, and the G+C content was 64.9%. There were 75 amino acid variation sites among 361 amino acids encoded by SLA-1 gene. The coding region of exon 2 and exon 3 had the highest polymorphism, and the degree of amino acid variation in exon 2 region was higher than that in exon 3 region. Among the 33 key amino acid sites that make up the six pockets of the peptide-binding groove, 11 of them were highly conserved between human and Rongchang pig, and 10 of the 19 key amino acid sites were consistent with β2-microglobulin. Only two sites, 225 (Thr/Ser) and 228 (Thr/Met), were different among the key amino acid sites that CD8 molecules bind to MHC. The analysis of homology and phylogenetic tree showed that SLA-1^*10:03 and SLA-1^*18:03 had the highest homology with human HLA-A^*02:01 and HLA-A^*11:01 alleles, respectively. Rongchang pig was closely related to Asian boar, Bama miniature pig, Rongshui miniature pig and other Asian pig breeds. The SLA-1 alleles of Chinese Rongchang pigs were successfully identified, and it was found to have highly rich polymorphism. The results of this study provided a genetic foundation for revealing the SLA genetic background of Rongchang pigs and conducting xenotransplantation research.

Key words: Rongchang pig, SLA-1 allele, identification, molecular characteristics

CLC Number:

S852.4

LIU Hongyi, LUO Tingyu, LI Changwen, YU Haibo, LU Xiaoye, CHEN Hongyan, XIA Changyou, GAO Caixia. Identification of SLA-1 Alleles and Analysis of Molecular Genetic Characteristics in Rongchang Pigs[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(7): 3064-3077.

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Identification of SLA-1 Alleles and Analysis of Molecular Genetic Characteristics in Rongchang Pigs

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