| 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.Attributed to the extremely array rhythm of rat germinal cells in seminiferous tubule, the method of DDRT-PCR combined with segmentation technique of the seminiferous tubule was adopted to isolate and identify the differentially expressed genes in spermatogenesis. Four segments i.e. II ~VI stages â…¦-â…§ stages â…¨~ â…« stages and â…©â…¢-â… stages were isolated under the anatomic microscope depending on the differential dioptres.The total RNA of these four segments were extracted and reverse transcripted to cDNA, and then were amplified with 3 single base-anchored polyT12N(N is C,G or T) primers matched with four upstream arbitrary primers. The products were electrophoresly segregated and auto-radiophotographed. 114 differential cDNA bands were picked out and 54 were confirmed to be of difference in different segments by reverse Northern dot-blot hybridyzation. 30 cDNA bands with distinct difference in expression quantity were selected to be cloned and sequenced ,and 20 novel ESTs were obtained by comparing the GenBank database and submitting the new sequences to GenBank.Based on the reverse Northern -blot hybridyzation results, 6 ESTs were selected to screen the cDNA library of rat testis, and three novel full-length cDNAs were obtained with the GenBank Accession Number as AF227439, AF146738, AF154849. The following are the characteristics of the three novel cDNA sequences, and the results of the two new genes' expression pattern and preliminary functional study.
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