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Expression of a novel pyridoxal kinase mRNA splice variant,PKH-T, in human testis

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Asian Journal of Andrology 2004年第2期6卷 83-91页

作者： xingFang zuo-minzhou lilu lan-lanyin jian-MinLi YinZhen HuiWang jia-haosha KeyLaboratoryofReproductiveMedicine NanjingMedicalUniversityNanjing210029China

Aim: To identify the genes specifically expressed in human adult and fetal testes and spermatozoa. Methods: A human testis cDNA microarray was established. Then mRNAs of human adult and fetal testis and spermatozoa were purified and probes were prepared by a reverse transcription reaction with mRNA as the template. The microarray was hybridized with probes of adult and fetal testes and spermatozoa. The nucleic acid sequences of differentially expressed genes were determined and homologies were searched in the databases of GenBank. Results: A novel human testis-specific gene, PKH-T, was identified by hybridizing adult and fetal testis and spermatozoa probes with a human testis cDNA microarray. The cDNA of PKH-T was 1 069 bp in length. The cDNA sequence of this clone was deposited in the Genbank (AY303972) and PKH-T was also determined as Interim GenSymbol (Unigene, HS.38041). PKH-T contained most PKH conserved motif. The 239 amino acid sequences deduced from the 719 bp open reading frame (ORF) had a homology with the gene PKH (U89606). PKH-T was specifically and strongly expressed in the testis. Comparison of the differential expressions of PKH and PKH-T in testes of different develop-mental stages indicated that PKH-T was expressed in the adult testis and spermatozoa, while PKH, in the adult, fetal and aged testes. PKH-T had no expression in the testis of Sertoli cell only and partially spermatogenic arrest patients. Conclusion: PKH-T is a gene highly expressed in adult human testis and spermatozoa. It may play an important role in spermatogenesis and could be related to male infertility. (Asian J Androl 2004 Jun; 6: 83-91)

关键词： testis-specific human pyridoxal kinase testis cDNA microarray spermatogenesis tissue distribution spermatogenic arrest ubiquitin

Identification of a novel testis-specific gene and its potential roles in testis development/spermatogenesis

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Asian Journal of Andrology 2005年第2期7卷 127-137页

作者： lan-lanyin jian-MinLi zuo-minzhou jia-haosha KeyLaboraryofReproductiveMedicine NanjingMedicalUniversityHangzhongRoadNanjing210029China

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 *** expressed genes were isolated and ***-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 ***:A novel testis-specific gene, NYD-SP5,was differentially expressed in fetal and adult *** 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 ***-PCR analysis revealed that NYD-SP5 was predominantly expressed in the testis but not in other 15 tissues *** 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 ***:NYD-SP5 is a newly found testis- specific gene with potential roles in testis development and spermatogenesis through a calmodulin-activated enzyme.

关键词： spermatogenesis testis calmodulin

Identification and characterization of cul-3b,a novel hominine CUL-3 transcript variant

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Asian Journal of Andrology 2005年第2期7卷 205-211页

作者： lilu zuo-MingZhou Xiao-YanHuang MinXu lan-lanyin HuiWang Zhi-YangXu jia-haosha KeyLaboratoryofReproductiveMedicine NanjingMedicalUniversityNanjing210029China

Aim:To identify genes related to the human testis development by substrate hybridization ***:A human testis cDNA microarray was constructed and hybridized with probes prepared from human adult and fetal testes and spermatozoa mRNAs by reverse transcription *** differentially expressed genes were sequenced. And a newly identified cullin-3 (CUL-3) transcript variant (designated cul-3b) was bio-informatically analyzed with an online GenBank ***-tissue reverse transcription polymerase chain reaction (RT-PCR) was used to deter- mine the tissue expression profile of ***:Cul-3b,a novel CUL-3 transcript variant,was *** expression level of cul-3b in adult testes was 3.79-fold higher than that in fetal ***-3b differed from cul-3 (including NM_003590 and AY337761) in the opening reading frame and had three internal ribosomal entry sites (IRESes) in the 5'-*** led to a 24 amino acid (aa) truncation at N-terminus of CUL-3b as compared with CUL-3 and a more motivated expression pattern of cul-3b under some strict ***,cul-3b expressed ubiquitously in human tissues according to multi-tissue ***:Cul-3b is a novel transcript variant of CUL-3,which may be important not only for the development of human testis but also for that of other organs.

关键词： alternative splicing CUL-3 DNA sequence human testis microarray