| Spermatogenesis is a series of highly ordered process which involves many genes expression under precise temporal and spatial regulation. The cloning and identification of these differentially expressed genes is of great value to delineate the mechanism of spermatogenesis.A cDNA, designated as rtSH3p13, was isolated from a rat testis cDNA library. It consisted of 1463 bp, Containing an open reading frame of 936 bp, coding a protein composed of 312 amino acids and assigned the GenBank accession number AF227439. The deduced rtSH3p13 protein is related to endophilins, members of the SH3p4/p8/p13 protein family isolated previously from a mouse brain cDNA expression library. The rtSH3p13 gene is expressed in the testis and brain, based on Northern blot analysis of various tissues. During postnatal development of the rat, rtSH3p13 is detected in the testis on day 18 and increases gradually thereafter to attain a maximum level on day 60. Using anti-rtSH3p13 anti-serum raised against the recombinant specific region of rtSH3p13, the antigen was immunolocalized in spermatids at steps 8-19 of spermiogenesis and found to be situated in the acrosome region by immuno-electron microscopy.Based on this background, we began our functional research of rtSH3p13 gene. GST "pull-down" assay and Co-immunoprecipitation method were used, and the results demonstrated that rtSH3p13 interacted with synaptojanin I, which is known to play a role in endocytosis. Then rtSH3p13 gene was cloned into pEGFP-N1, and the recombinant plasmid was transfected into CHO cells to express the fusion protein of rtSH3p13-GFP. The expression of the protein was confirmed by Western Blot analysis as well as cellular fluorescence assay. To further explore the physiological function of the rtSH3p13 protein, the...
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