这项研究的目的是确定有效的遗传标记的抗原加工相关转运蛋白1(TAP1),α(1,2)岩藻糖基转移酶1(FUT1),天然抗性相关巨噬细胞蛋白1(NRAMP1),当地猪品种的粘蛋白4(MUC4)和粘蛋白13(MUC13)腹泻抗性基因,即上海白猪,枫井猪,Shawutou猪,梅山猪和浦东白猪,为上海地方猪品种资源的鉴定提供参考。应用聚合酶链反应-限制性片段长度多态性(PCR-RFLR)和序列测序分析上述基因的多态性,并结合一些免疫因素探讨对上海地方猪品种免疫的影响。结果表明,在5个猪品种中,TAP1和MUC4基因均具有抗腹泻基因型GG,在浦东白猪中检测到FUT1基因的AG和GG基因型,在梅山猪中检测到NRAMP1基因的AA抗腹泻基因,在浦东白猪中检测到AB型NRAMP1基因,MUC13基因的抗腹泻基因型GG仅在上海白猪中检测到。MUC13抗腹泻基因型GG仅在上海白猪中检测到。TAP1基因在上海白猪中呈中度多态性,枫井猪,Shawutou猪,梅山猪和浦东白猪,其中上海白猪和沙乌头猪的TAP1不满足哈代-温伯格均衡。浦东白猪FUT1基因处于低多态性状态。眉山猪和浦东白猪的NRAMP1处于中度多态性状态,不满足哈代-温伯格均衡。上海白猪和浦东白猪的MUC4基因处于低多态性状态,风井猪和沙乌头猪的MUC4基因处于中度多态性状态,枫井猪和浦东白猪的MUC4基因不满足Hardy-Weinberg平衡。上海白猪和浦东白猪的MUC13基因处于中度多态性状态。梅山猪IL-2、IL-10、IgG和TNF-α水平较高,浦东白猪的IL-12水平高于其他猪。上海白猪MUC13基因AA基因型的白细胞介素12(IL-12)水平明显高于AG基因型。风雨猪TAP1基因AA基因型的肿瘤坏死因子α(TNF-α)指标明显高于GG和AG基因型。Shawutou猪TAP1基因AG基因型中IL-12的指标明显高于GG基因型。梅山猪NRAMP1基因AA基因型的TNF-α水平明显高于AB基因型。FUT1基因AG型的IL-2水平明显高于GG型的浦东白猪,MUC4基因AA型的IL-2水平明显高于AG型,MUC13基因GG型IgG水平明显高于AG型。本研究结果对指导上海白猪的止泻选育和分子筛选具有重要意义,枫井猪,Shawutou猪,梅山猪和浦东白猪,为今后上海各地方猪种的抗腹泻育种奠定了基础。
The aim of this study was to identify effective genetic markers for the Antigen Processing Associated Transporter 1 (TAP1), α (1,2) Fucosyltransferase 1 (FUT1), Natural Resistance Associated Macrophage Protein 1 (NRAMP1), Mucin 4 (MUC4) and Mucin 13 (MUC13) diarrhea-resistance genes in the local pig breeds, namely Shanghai white pigs, Fengjing pigs, Shawutou pigs, Meishan pigs and Pudong white pigs, to provide a reference for the characterization of local pig breed resources in Shanghai. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLR) and sequence sequencing were applied to analyze the polymorphisms of the above genes and to explore the effects on the immunity of Shanghai local pig breeds in conjunction with some immunity factors. The results showed that both TAP1 and MUC4 genes had antidiarrheal genotype GG in the five pig breeds, AG and GG genotypes of the FUT1 gene were detected in Pudong white pigs, AA antidiarrheal genes of the NRAMP1 gene were detected in Meishan pigs, the AB type of the NRAMP1 gene was detected in Pudong white pigs, and antidiarrheal genotype GG of the MUC13 gene was only detected in Shanghai white pigs. The MUC13 antidiarrhea genotype GG was only detected in Shanghai white pigs. The TAP1 gene was moderately polymorphic in Shanghai white pigs, Fengjing pigs, Shawutou pigs, Meishan pigs and Pudong white pigs, among which TAP1 in Shanghai white pigs and Shawutou pigs did not satisfy the Hardy-Weinberg equilibrium. The FUT1 gene of Pudong white pigs was in a state of low polymorphism. NRAMP1 of Meishan pigs and Pudong white pigs was in a state of moderate polymorphism, which did not satisfy the Hardy-Weinberg equilibrium. The MUC4 genes of Shanghai white pigs and Pudong white pigs were in a state of low polymorphism, and the MUC4 genes of Fengjing pigs and Shawutou pigs were in a state of moderate polymorphism, and the MUC4 genes of Fengjing pigs and Pudong white pigs did not satisfy the Hardy-Weinberg equilibrium. The MUC13 gene of Shanghai white pigs and Pudong white pigs was in a state of moderate polymorphism. Meishan pigs had higher levels of IL-2, IL-10, IgG and TNF-α, and Pudong white pigs had higher levels of IL-12 than the other pigs. The level of interleukin 12 (IL-12) was significantly higher in the AA genotype of the MUC13 gene of Shanghai white pigs than in the AG genotype. The indicator of tumor necrosis factor alpha (TNF-α) in the AA genotype of the TAP1 gene of Fengjing pigs was significantly higher than that of the GG and AG genotypes. The indicator of IL-12 in the AG genotype of the Shawutou pig TAP1 gene was significantly higher than that of the GG genotype. The level of TNF-α in the AA genotype of the NRAMP1 gene of Meishan pigs was markedly higher than that of the AB genotype. The IL-2 level of the AG type of the FUT1 gene was obviously higher than that of the GG type of Pudong white pigs, the IL-2 level of the AA type of the MUC4 gene was dramatically higher than that of the AG type, and the IgG level of the GG type of the MUC13 gene was apparently higher than that of the AG type. The results of this study are of great significance in guiding the antidiarrhea breeding and molecular selection of Shanghai white pigs, Fengjing pigs, Shawutou pigs, Meishan pigs and Pudong white pigs and laying the foundation for future antidiarrhea breeding of various local pig breeds in Shanghai.