Expression Of Inhibin α Subunit In Male Rat And Construction Of Its Promoter Expression Vector

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.