LncRNA调控E.coli型奶牛乳房炎的分子机制

E.coli是引起奶牛乳房炎的常见病原体。研究表明，lncRNA可调节人和小鼠的炎症，而对E.coli型奶牛乳房炎的分子调控机制尚不清楚。因此，本课题拟对前期诱导的E.coli型乳房炎奶牛的乳腺组织（健康奶牛为对照组）进行lncRNA测序，结合前期对同一批组织测定的miRNA和转录组数据，进行多组学联合分析，筛选出与E.coli型乳房炎相关的差异表达lncRNA。通过组织/细胞表达分析、超表达和干扰等方法，研究差异表达lncRNA对乳腺上皮细胞炎症反应的影响，以确定具有重要功能的1-3个关键lncRNA。同时采用亚细胞定位、双荧光素酶报告基因系统、ChIRP-seq、RNA pull-down、qPCR、WB、RIP和流式细胞等技术，系统揭示关键lncRNA调控E.coli型乳房炎的分子机制，为解析奶牛乳房炎的分子调控网络奠定基础，为培育抗乳房炎奶牛新品种提供理论依据。

E. coli is a common pathogen of causing cow mastitis. The previous studies showed that lncRNA can regulate the inflammation of human and mice, but the molecular mechanism of regulating cow E.coli-type mastitis is not clear. Therefore, this project intends to perform the lncRNA sequencing of the E.coli-induced mammary tissues (healthy cow as control group). Combined with the data of miRNA and transcriptome sequencing in the same mammary tissues mentioned above, integrated analysis of multiple omics will be used to screen the differentially expressed lncRNA related to E.coli-type mastitis. The methods of tissue / cell expression analysis, gene overexpression and interference will be used to clarify the effects of differentially expressed lncRNA on the inflammatory response of mammary epithelial cells to identify 1-3 lncRNAs with important functions. Meanwhile, lots of methods, including subcellular localization of lncRNA, double luciferase reporter gene assay, ChIRP-seq, Western Blot, RNA pull-down, qPCR, RIP and flow cytometry, will be used to systematically investigate the molecular mechanism of lncRNA regulating E.coli-type mastitis. The results will lay a foundation for clarifying the molecular regulatory network of cow mastitis and provide a theoretical basis for the breeding of dairy cattle resistant to mastitis.