[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αβT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。
Genome-wide Association Study of Anti-brucellosis Traits Based on cELISA Detection in Sheep
[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.
sheepbrucellosiscompetitive enzyme-linked immunosorbent assay(cELISA)genome-wide association study(GWAS)
万方数据
维普
