【作者】 黄翔；

【导师】 熊承良；

【作者基本信息】 华中科技大学 ， 妇产科学， 2009， 硕士

【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。更多还原

【Abstract】 Objective: To examine the expression of Inhibinαprotein and the relative expression levels of its mRNA in various tissues.Methods: Adult male SD rats were killed immediately after anesthesia with 10% chloral hydrate.The testis, epididymis, kidney, adrenal and liver were isolated, The immunohistochemistry and fluorescence quantitative PCR technology was performed to detect the Inhibinαprotein and mRNA in the rat tissues.Results: Inhibinαprotein was expressed in testis,epididymis, kidney adrenal gland and pituitary but not in liver. The relative expression levels of inhibinαmRNA were as follows: kidney> adrenal> testis> epididymis.Conclusion: Both qualitative and quantitative detection of Inhibinαsubunit in this experiment suggest that the Inhibinαsubunit is not only expressed in tissues of reproductive system but also in other tissues. Objective: To construct eukaryotic expression vector of rat Inhibinαgene promoter region respectively after fragmental cloning, in order to prepare for selecting its core promoter.Methods: Papers and network resources had been used for the prediction and the bioinformatic analysis of Inhibinαpromoter region.Gene recombinant technology was used to clone the fragmental promoter region into the eukaryotic expression vector ------pEGFP-N1, then the sequences were verified.Results: 395bp, 606bp and 775bp fragments of the rat Inhibinαgene promoter region were cloned successfully, moreover, the eukaryotic expression vectors containing this three fragments were constructed respectively. Conclusion: The predicted rat Inhibinαgene promoter region is successfully cloned and the eukaryotic expression vectors containing the fragmental promoter are successfully constructed.更多还原