基于cELISA检测的绵羊抗布鲁氏菌病性状全基因组关联分析

doi:10.16431/j.cnki.1671-7236.2024.10.001

摘要/Abstract

摘要： 【目的】挖掘与绵羊布鲁氏菌病抗性或易感性相关的基因，以期为绵羊布鲁氏菌病抗病育种提供分子标记。【方法】以自然感染布鲁氏菌病并且未注射疫苗的绵羊为研究对象，采集50只绵羊的血液样本进行竞争酶联免疫吸附试验(competitive enzyme-linked immunosorbent assay，cELISA)。对绵羊血液DNA进行全基因组重测序，以cELISA结果为数量性状表型，利用GEMMA软件进行全基因组关联分析。使用clusterProfiler软件包对显著基因进行功能富集分析。【结果】全基因组关联分析筛选到Top 1 000的位点共注释到56个基因，GO功能富集结果显示，注释基因主要富集在β选择和αβ T细胞增殖和分化等条目；KEGG通路富集分析结果显示，注释基因主要富集在Th1/Th2/Th17细胞分化和自然杀伤细胞介导的细胞毒性等通路，主要涉及γ-干扰素受体1(interferon gamma receptor 1，IFNGR1)、活化T细胞核因子2(nuclear factor of activated T cells 2，NFATC2)、NF-κB抑制因子β(NF-κB inhibitor beta，NFKBIB)、脾酪氨酸激酶(spleen associated tyrosine kinase，SYK)、转化生长因子β受体2(transforming growth factor beta receptor 2，TGFBR2)和T细胞受体相关蛋白激酶70的Zeta链(Zeta chain of T cell receptor associated protein kinase 70，ZAP70)共6个基因，筛选到的条目、通路和基因与布鲁氏菌的感染或宿主的抗感染密切相关。【结论】本研究筛选到6个与绵羊布鲁氏菌病相关的基因，为进一步细胞或个体水平的功能验证奠定了基础，为绵羊布鲁氏菌病抗病育种分子标记提供了参考。

关键词: 绵羊; 布鲁氏菌病; 竞争酶联免疫吸附试验(cELISA); 全基因组关联分析(GWAS)

Abstract: 【Objective】 The aim of this study was to explore genes associated with brucellosis resistance or susceptibility in sheep，in order to provide molecular markers for brucellosis resistance breeding in sheep. 【Method】 In this study，sheep naturally infected with brucellosis and not vaccinated were used as research subjects，and blood samples were collected for competitive enzyme-linked immunosorbent assay (cELISA). The whole-genome resequencing was conducted on the blood DNA samples. The results of cELISA were used as quantitative trait phenotypes，and GEMMA software was used for genome-wide association study. The clusterProfiler package was used to perform enrichment analysis on significant genes. 【Result】 A total of 56 genes were annotated from the Top 1 000 loci selected by genome-wide association study. GO functional enrichment results showed that annotated genes were mainly concentrated in β-selection and αβ T cell proliferation and differentiation. KEGG pathway enrichment analysis showed that annotated genes were mainly concentrated in Th1/Th2/Th17 cell differentiation and natural killer cell-mediated cytotoxicity. It mainly involved 6 genes，including interferon gamma receptor 1 (IFNGR1)，nuclear factor of activated T cells 2 (NFATC2)，NF-κB inhibitor beta (NFKBIB)，spleen associated tyrosine kinase (SYK)，transforming growth factor beta receptor 2 (TGFBR2)，and Zeta chain of T cell receptor associated protein kinase 70 (ZAP70). The screened items，pathways，and genes were closely related to Brucella infection or the host’s anti-infection response. 【Conclusion】 6 genes related to sheep brucellosis were screened，which laid a foundation for further functional verification at the cellular or individual level，and provided a reference for molecular markers for breeding sheep brucellosis resistance.

Key words: sheep; brucellosis; competitive enzyme-linked immunosorbent assay (cELISA); genome-wide association study (GWAS)

中图分类号:

S813.2

武上杰, 李佳佳, 蒋琳, 马月辉, 丁家波, 何晓红. 基于cELISA检测的绵羊抗布鲁氏菌病性状全基因组关联分析[J]. 中国畜牧兽医, 2024, 51(10): 4191-4199.

WU Shangjie, LI Jiajia, JIANG Lin, MA Yuehui, DING Jiabo, HE Xiaohong. Genome-wide Association Study of Anti-brucellosis Traits Based on cELISA Detection in Sheep[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(10): 4191-4199.

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