盘羊及其杂交羊SPLUNC1特性研究及抗肺炎支原体差异表达基因的筛选

项目名称： 盘羊及其杂交羊SPLUNC1特性研究及抗肺炎支原体差异表达基因的筛选

项目编号： No.31460686

项目类型： 地区科学基金项目

立项/批准年度： 2015

项目学科： 农业科学

项目作者： 孙延鸣

作者单位： 石河子大学

项目金额： 50万元

中文摘要： 盘羊与巴什拜羊杂交为种间杂交，其杂交后代对肺炎支原体感染抗性减弱，并造成在生长发育早期大量死亡。短腭、肺及鼻咽上皮克隆1（SPLUNC1）为一分泌性蛋白，在促进肺部先天防御肺炎支原体感染过程中发挥了重要作用。本研究以新疆野生盘羊、巴什拜羊及其杂交后代为研究对象，以支原体肺炎为疾病模型，从羊的SPLUNC1基因入手，通过人工授精、基因克隆、测序、基因表达、real-time PCR、细胞培养、电镜观察，流式细胞、人工感染、ELISA等研究手段，进行比较基因组学分析，比较以上不同品种羊SPLUNC1蛋白的功能，以期从基因结构、基因表达水平、蛋白功能、实际抗病等多个角度去揭示SPLUNC1在盘羊杂交后代对肺炎支原体抗性减弱而死亡中所起到的作用。同时运用数字基因表达谱技术，筛选不同品种羊抗肺炎支原体差异表达基因，为进一步研究杂交羊易感支原体肺炎的分子作用机制奠定基础。

中文关键词： 盘羊；巴什拜羊；短腭；肺及鼻咽上皮克隆1；肺炎支原体；差异表达基因

英文摘要： The hybridization of wild argali and bashbay sheep is interspecific hybridization. The resistance of Mycoplasma pneumoniae (Mp) infection in the hybrid offspring is weakened, which arosed a lot of death in these mules in early growth. Short palate，lung，and nasal epithelium clone 1 (SPLUNC1) serves as a secretory protein，which play an important role in promoting lung innate defense against Mp infection. In this study, Xinjiang wild argali and bashbay sheep and their hybrid sheep are objects. Mycoplasma pneumonia of sheep is the disease model. Study on SPLUNC1 gene by using techniques such as artificial insemination, gene clone, gene sequencing, gene expression, real-time PCR, cell culture, electron microscope, flow cytometry, artificial infection and ELISA. Comparative genomics in argali, bashbay sheep and their hybrid sheep will be analyzed.Function of SPLUNC1 protein from different species sheep will be compared. Effects of SPLUNC1 in these argali hybrids that reduced Mp infection resistances as well as death will be revealed from genetic structure, gene expression level, protein function, actual disease resistance. Meanwhile, differentially expressed genes of anti-Mp from different species sheep will be screened by Digital Gene Expression Profiling technology. It can also provide a theoretical basis for further research the molecular mechanism of susceptible Mp in the hybrid sheep.

英文关键词： argali;bashbay sheep;short palate;lung and nasal epithelium clone 1;mycoplasma pneumoniae;differentially expressed genes