Cloning of CDC25A gene differentially expressed in testis tissues of yak and cattle-yak
[Objective]The present study aimed to investigate the expression differences of the CDC25A gene in the testicular tissues and spermatogenic cells of yak and cattle-yak,as well as the similarities and differences in the two gene sequences.[Method]The yak and cattle-yak were selected as experimental animals,and the expression levels of CDC25A gene in testis tissues and spermatogenic cells of yak and cat-tle-yak were detected by real-time fluorescence quantitative PCR,and the CDC25A cDNAs of yak and cattle-yak testis tissues were amplified and cloned by RT-PCR method.The structures and functions of these two were analyzed using bioinformatics tools.[Result]Immunofluores-cence identification revealed positive expression of the germ cell marker protein(DDX4)in the suspension cells of yaks and cattle-yak,indi-cating that this cell population consisted of spermatogenic cells.The expression of CDC25A in testis tissues and spermatogenic cells of cattle-yak was extremely significantly lower than that of yak(P＜0.01).The CDC25A gene sequences of yak and cattle-yak were obtained by RT-PCR,with a length of 1749 and 1744 bp,respectively.Both of the CDC25A CDS regions were 1434 bp,encoding 477 amino acids.The phylo-genetic tree constructed by using the neighbor-joining method indicated that the CDC25A genes of yak and cattle-yak had the closest relation-ship with the CDC25A of wild yak.The CDC25A protein contained a total negative charge residue(Asp+Glu)charge of 73 and a total posi-tive charge residue(Arg+Lys)charge of 66,had no transmembrane structure and signal peptide structure,and were unstable hydrophilic ex-tracellular proteins.Subcellular localization results showed that it was primarily located in the nucleus.[Conclusion]In the study,the expres-sion levels of CDC25A gene were significantly different in the testis tissues and spermatogenic cells of yaks and cattle-yak,and the gene se-quences were successfully obtained.However,CDC25A in cattle-yak was basically consistent with that in maternal yaks,with no significant differences,indicating that the obstruction of spermatogenesis was not due to gene mutation but associated with the reduced expression of CDC25A.These results can provide a reference for further studies on the mechanism of CDC25A participating in spermatogenic cell prolifera-tion of cattle-yak,and also provide a new idea for the study of male sterility of cattle-yak.
万方数据
维普
