雄性大鼠Inhibinα亚基的表达及其启动子表达载体的构建

发布时间：2018-06-21 07:54

本文选题：大鼠 + Inhibinα蛋白　；参考：《华中科技大学》2009年硕士论文

【摘要】：目的:确定雄性大鼠Inhibinα亚基的mRNA及蛋白在各种组织中有无表达及相对表达量。方法:选用成年SD雄性大鼠,10%水合氯醛麻醉后处死,立即取睾丸、附睾、肾、肾上腺及肝组织,采用免疫组化法和荧光定量PCR方法分别检测Inhibinα蛋白和mRNA在大鼠各组织中的表达情况。结果:Inhibinα蛋白在睾丸、附睾、肾、肾上腺组织中均有表达,在肝脏组织中无表达。InhibinαmRNA在各组织中的表达相对量依次为:肾肾上腺睾丸附睾。结论:本实验从定性和定量两方面检测了Inhibinα亚基的表达情况,结果提示Inhibinα亚基不仅仅在生殖系统有表达,在其它系统也有表达。目的:将大鼠Inhibinα基因的启动子区分段克隆后分别构建真核表达载体,为进一步筛选出其核心启动子做好准备。方法:利用已有文献及网络资源进行Inhibinα启动子区的预测及生物信息学分析,用基因重组技术将预测的启动子区分段克隆到真核表达载体pEGFP-N1中,并对其测序验证。结果:成功克隆了大鼠Inhibinα基因的启动子区的395bp、606bp、775bp大小的片段,并分别构建了这三种片段的真核表达载体。结论:对预测的大鼠Inhibinα基因的启动子区分段克隆,并成功构建了含有这些克隆片段的真核表达载体。
[Abstract]:Aim: to determine the mRNA and protein expression of Inhibin 伪 subunit in various tissues of male rats. Methods: adult SD male rats were killed after anaesthesia with 10% chloral hydrate. Testis, epididymis, adrenal gland and liver tissues were collected immediately. The expression of Inhibin 伪 protein and mRNA in rat tissues was detected by immunohistochemistry and fluorescence quantitative PCR, respectively. Results the percent Inhibin 伪 protein was expressed in testis, epididymis and adrenal gland. Conclusion: the expression of Inhibin 伪 subunit was detected qualitatively and quantitatively in this experiment. The results suggest that the expression of Inhibin 伪 subunit is not only expressed in reproductive system, but also in other systems. Aim: to construct the eukaryotic expression vector of rat Inhibin 伪 gene promoter after cloning, so as to prepare for the further screening of its core promoter. Methods: the predicted promoter differentiation segment was cloned into the eukaryotic expression vector pEGFP-N1 by gene recombination technique, and was sequenced and verified by using the existing literature and network resources to predict the promoter region of Inhibin 伪 and bioinformatics analysis. Results: the promoter region of rat Inhibin 伪 gene was successfully cloned and cloned, and the eukaryotic expression vectors of these three fragments were constructed. Conclusion: the promoter differentiation cloning of the predicted rat inhibin 伪 gene was carried out and the eukaryotic expression vector containing these fragments was successfully constructed.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R346

【参考文献】