Aim:To identify and characterize a novel gene with potential roles in testis development and spermatogenesis. Methods:A cDNA microarray was constructed from a human testis large insert cDNA library and hybridized with...Aim:To identify and characterize a novel gene with potential roles in testis development and spermatogenesis. Methods:A cDNA microarray was constructed from a human testis large insert cDNA library and hybridized with probes of human or mouse adult and fetal testes.Differentially expressed genes were isolated and sequenced.RT-PCR was used to test the tissue distribution of the genes of interest and in situ hybridization was performed to localize the gene expression in the mouse testis.A range of bioinformatical programs including Gene Runner,SMART,NCBI Blast and Emboss CpGPlot were used to characterize the new gene's feature.Results:A novel testis-specific gene, NYD-SP5,was differentially expressed in fetal and adult testes.The deduced protein structure of NYD-SP5 was found to contain an IQ motif (a short calmodulin-binding motif containing conserved lie and Gin residues),a Carbam- ate kinase-like domain,a Zn-dependent exopeptidase domain and a lactate dehydrogenase (LDH) C-terminal-like domain.RT-PCR analysis revealed that NYD-SP5 was predominantly expressed in the testis but not in other 15 tissues examined.In situ hybridization and RT-PCR examinations revealed that the expression of NYD-SP5 was confined in the male germ cell but not present in the somatic cell in the testes.Conclusion:NYD-SP5 is a newly found testis- specific gene with potential roles in testis development and spermatogenesis through a calmodulin-activated enzyme.展开更多
文摘Aim:To identify and characterize a novel gene with potential roles in testis development and spermatogenesis. Methods:A cDNA microarray was constructed from a human testis large insert cDNA library and hybridized with probes of human or mouse adult and fetal testes.Differentially expressed genes were isolated and sequenced.RT-PCR was used to test the tissue distribution of the genes of interest and in situ hybridization was performed to localize the gene expression in the mouse testis.A range of bioinformatical programs including Gene Runner,SMART,NCBI Blast and Emboss CpGPlot were used to characterize the new gene's feature.Results:A novel testis-specific gene, NYD-SP5,was differentially expressed in fetal and adult testes.The deduced protein structure of NYD-SP5 was found to contain an IQ motif (a short calmodulin-binding motif containing conserved lie and Gin residues),a Carbam- ate kinase-like domain,a Zn-dependent exopeptidase domain and a lactate dehydrogenase (LDH) C-terminal-like domain.RT-PCR analysis revealed that NYD-SP5 was predominantly expressed in the testis but not in other 15 tissues examined.In situ hybridization and RT-PCR examinations revealed that the expression of NYD-SP5 was confined in the male germ cell but not present in the somatic cell in the testes.Conclusion:NYD-SP5 is a newly found testis- specific gene with potential roles in testis development and spermatogenesis through a calmodulin-activated enzyme.
