Gene Cloning And Expression Of Mouse Testis-Specific Serine/Threonine Kinase2

The testis-specific serine/threonine kinase family is the kinases that exclusively express in male germ cells and/or in the testis. The restricted tissue and time expression pattern of Tssks suggests that Tssk(s) have an important role(s) in the regulation of spermatogenesis and/or sperm function.In this experiment, the full length cDNA of mouse Tssk2has been cloned via RT-PCR strategy, then the prokaryotic expression system of mouse Tssk2has been constructed by subcloning Tssk2’s ORF into pGEX-5x-1vector and transforming to E.coli BL21. After western blot analysis, the extracted, purified protein product, weighing about65kDa, was proved to be a Tssk2fusion protein with GST tag.In addition, the expression and subcellular localization of Tssk2in mouse and human sperm have been examined by Western blot and immunohistochemistry. The results show that Tssk2is present in mouse and human sperm, Tssk2mainly distributes in post-acrosomal region and equatorial region in mouse and human sperm, respectively.