基于基因组学技术鉴定奶牛副结核病易感/抗性基因

项目名称： 基于基因组学技术鉴定奶牛副结核病易感/抗性基因

项目编号： No.31472065

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 畜牧学与草地科学

项目作者： 孙东晓

作者单位： 中国农业大学

项目金额： 88万元

中文摘要： 旨在以北京地区中国荷斯坦牛为试验群体，鉴定奶牛副结核病易感/抗性基因，研究内容包括：1）采用ELISA方法检测奶牛血清副结核菌抗体水平，基于牛50K SNP芯片，采用基于case-control设计的全基因组关联分析（GWAS）检测与奶牛血清副结核菌抗体水平病显著关联的SNP位点，通过生物信息学分析，鉴定奶牛副结核病易感/抗性基因并功能注释；2）采用基于第二代高通量测序的RNA-seq、miRNA-seq、CHIP-seq技术，针对副结核菌感染且临床发病、感染但未发病、健康的奶牛个体空肠组织进行转录组、microRNA、转录因子和组蛋白修饰分析，鉴定差异表达基因及调控通路；3）综合上述4个层次的数据分析结果，筛选易感/抗性候选基因并进行大群验证及细胞水平功能分析，最终鉴定奶牛副结核病易感/抗性关键基因和致因突变，为实施奶牛副结核病抗性标记辅助预选青年公牛和母牛提供有效的分子信息。

中文关键词： 荷斯坦奶牛；全基因组关联分析；二代测序；牛副结核分枝杆菌；易感/抗性基因

英文摘要： The purpose of this study is aimed to detect the serum antibody levle of Mycobacterium avium subsp. Paratuberculosis (MAP) with ELISA kit and identify genes affecting susceptibility/resistence to MAP in the Holstein population in Beijing region.1) Based on 50K SNP chip, genome-wide association analysis (GWAS) will be conducted for susceptibility/resistenc to MAP with case-control design to identify genome-wide and chromosome-wide significant SNPs. Through further bioinformation analysis, candidate genes for such trait will be mined and function annotiated; 2) Using RNA-seq，miRNA-seq,CHIP-seq technologies which are based on the next generation high- throughput sequencing technique, jejunum tissue samples will be selected from 9 Holstein cows those were infected with MAP and showed clinical symptom, infected with MAP but no symptom, and health, respectively, with 3 cows in each group, and conducted transcriptome, miRNA, transcript factor and histone modification analysis to identify the significantly expressed genes among 3 groups and regulatory pathways; 3)Corresponding, integrated the above results, we will confirm the genetic effects of such candidate key genes on susceptibility/resistence to MAP in the independant Holstein population, and then investigate the biological functions of the candidates with large genetic effects based on jejunum cell line in order to determine the ture key genes and causal mutations, so that marker-assisted selection program on resistance to MAP will be put into effect for Holstein bulls and cows.

英文关键词： Holstein;GWAS;next-generation sequencing;M.paratuberculosis;susceptibility/resistence gene