Evolutionary evidence suggests that Sox3, a member of the high-mobility-group(HMG) family of transcription factors, is an ancestral precursor of Sry and is involved in sex determination similar to Sry. However, there ...Evolutionary evidence suggests that Sox3, a member of the high-mobility-group(HMG) family of transcription factors, is an ancestral precursor of Sry and is involved in sex determination similar to Sry. However, there is limited information regarding the SOX3 gene of the black rockfish(Sebastes schlegeli). In this study, we first isolated SOX3 gene from the gonads of S. schlegeli by homology cloning. The full-length of S. schlegeli SOX3(SsSOX3) c DNA was 1386 bp, comprising a 906-bp open reading frame, which encodes a peptide showing 93.6% and 93.9% homology with the Sox3 proteins of Epinephelus coioides and Oryzias latipe, respectively. Comparison of the cDNA sequence of the Ss SOX3 gene with the corresponding genomic DNA fragment revealed that the SsSOX3 gene consists of a single exon. Phylogenetic analysis demonstrated the evolutionary relationship of Ss SOX3 with other known SOXB1 genes in fish and tetrapods. The promoter region contains binding sites of several transcriptional factors that might participate in the regulation of Ss SOX3 expression. Quantitative real-time PCR analysis indicated that SsSOX3 was expressed in all the investigated larval developmental stages from 1 to 35 days after birth and the level of expression gradually decreased as the development proceeded. SsSOX3 exhibited sexually dimorphic expression in adult gonads, with high expression in the ovary but low expression in the testis. In situ hybridization revealed that SsSOX3 was strongly expressed in oocytes and follicular cells of ovaries but slightly expressed in germ cells of testicular tissues. Therefore, this study suggests that Ss SOX3 plays an important role in oogenesis and ovary differentiation in S. schlegeli.展开更多
Sex related FTZ-F1 is a transcriptional factor regulating the expression of fushi tarazu (a member of the orphan nuclear receptors) gene. In this study, FTZ-F1 gene (FTZ-F1) was isolated from the testis of black rockf...Sex related FTZ-F1 is a transcriptional factor regulating the expression of fushi tarazu (a member of the orphan nuclear receptors) gene. In this study, FTZ-F1 gene (FTZ-F1) was isolated from the testis of black rockfish (Sebastes schlegeli) by homology cloning. The full-length cDNA of S. schlegeli FTZ-F1 (ssFTZ-F1) contained a 232bp 5′UTR, a 1449bp ORF encoding FTZ-F1 (482 amino acid residules in length) with an estimated molecular weight of 5.4kD and a 105bp 3′UTR. Sequence, tissue distribution and phylogenic analysis showed that ssFTZ-F1 belonged to FTZ group, holding highly conserved regions including Ⅰ, Ⅱ and Ⅲ FTZ-F1 boxes and an AF-2 hexamer. Relatively high expression was observed at different larva stages. In juveniles (105 days old), the transcript of ssFTZ-F1 can be detected in all tissues and the abuncance of the gene transcript in testis, ovary, spleen and brain was higher than that in other tissues. In mature fish, the abundance of gene transcript was higher in testis, ovary, spleen and brain than that in liver (trace amount), and the gene was not transcribed in other tissues. The highest abundance of gene transcript was always observed in gonads of both juvenile and mature fish. In addition, the abundance of gene transcript in male tissues were higher than that in female tissue counterparts (P<0.05).展开更多
Cold-inducible RNA-binding protein(CIRP) is a kind of RNA binding proteins that plays important roles in many physiological processes. The CIRP has been widely studied in mammals and amphibians since it was first clon...Cold-inducible RNA-binding protein(CIRP) is a kind of RNA binding proteins that plays important roles in many physiological processes. The CIRP has been widely studied in mammals and amphibians since it was first cloned from mammals. On the contrary, there are little reports in teleosts. In this study, the Po CIRP gene of the Japanese flounder was cloned and sequenced. The genomic sequence consists of seven exons and six introns. The putative Po CIRP protein of flounder was 198 amino acid residues long containing the RNA recognition motif(RRM). Phylogenetic analysis showed that the flounder Po CIRP is highly conserved with other teleost CIRPs. The 5' flanking sequence was cloned by genome walking and many transcription factor binding sites were identified. There is a Cp Gs region located in promoter and exon I region and the methylation state is low. Quantitative real-time PCR analysis uncovered that Po CIRP gene was widely expressed in adult tissues with the highest expression level in the ovary. The m RNA of the Po CIRP was maternally deposited and the expression level of the gene was regulated up during the gastrula and neurula stages. In order to gain the information how the protein interacts with m RNA, we performed the modeling of the 3D structure of the flounder Po CIRP. The results showed a cleft existing the surface of the molecular. Taken together, the results indicate that the CIRP is a multifunctional molecular in teleosts and the findings about the structure provide valuable information for understanding the basis of this protein's function.展开更多
基金supported by the National Natural Science Foundation of China (No. 31372511)
文摘Evolutionary evidence suggests that Sox3, a member of the high-mobility-group(HMG) family of transcription factors, is an ancestral precursor of Sry and is involved in sex determination similar to Sry. However, there is limited information regarding the SOX3 gene of the black rockfish(Sebastes schlegeli). In this study, we first isolated SOX3 gene from the gonads of S. schlegeli by homology cloning. The full-length of S. schlegeli SOX3(SsSOX3) c DNA was 1386 bp, comprising a 906-bp open reading frame, which encodes a peptide showing 93.6% and 93.9% homology with the Sox3 proteins of Epinephelus coioides and Oryzias latipe, respectively. Comparison of the cDNA sequence of the Ss SOX3 gene with the corresponding genomic DNA fragment revealed that the SsSOX3 gene consists of a single exon. Phylogenetic analysis demonstrated the evolutionary relationship of Ss SOX3 with other known SOXB1 genes in fish and tetrapods. The promoter region contains binding sites of several transcriptional factors that might participate in the regulation of Ss SOX3 expression. Quantitative real-time PCR analysis indicated that SsSOX3 was expressed in all the investigated larval developmental stages from 1 to 35 days after birth and the level of expression gradually decreased as the development proceeded. SsSOX3 exhibited sexually dimorphic expression in adult gonads, with high expression in the ovary but low expression in the testis. In situ hybridization revealed that SsSOX3 was strongly expressed in oocytes and follicular cells of ovaries but slightly expressed in germ cells of testicular tissues. Therefore, this study suggests that Ss SOX3 plays an important role in oogenesis and ovary differentiation in S. schlegeli.
基金supported by the National High-Tech Research and Development Program (2012AA10A402)the National Natural Science Foundation of China(31172385)
文摘Sex related FTZ-F1 is a transcriptional factor regulating the expression of fushi tarazu (a member of the orphan nuclear receptors) gene. In this study, FTZ-F1 gene (FTZ-F1) was isolated from the testis of black rockfish (Sebastes schlegeli) by homology cloning. The full-length cDNA of S. schlegeli FTZ-F1 (ssFTZ-F1) contained a 232bp 5′UTR, a 1449bp ORF encoding FTZ-F1 (482 amino acid residules in length) with an estimated molecular weight of 5.4kD and a 105bp 3′UTR. Sequence, tissue distribution and phylogenic analysis showed that ssFTZ-F1 belonged to FTZ group, holding highly conserved regions including Ⅰ, Ⅱ and Ⅲ FTZ-F1 boxes and an AF-2 hexamer. Relatively high expression was observed at different larva stages. In juveniles (105 days old), the transcript of ssFTZ-F1 can be detected in all tissues and the abuncance of the gene transcript in testis, ovary, spleen and brain was higher than that in other tissues. In mature fish, the abundance of gene transcript was higher in testis, ovary, spleen and brain than that in liver (trace amount), and the gene was not transcribed in other tissues. The highest abundance of gene transcript was always observed in gonads of both juvenile and mature fish. In addition, the abundance of gene transcript in male tissues were higher than that in female tissue counterparts (P<0.05).
基金supported by the National High Technology R&D Program of China (2012AA10A402)the National Natural Science Foundation of China (31172385)
文摘Cold-inducible RNA-binding protein(CIRP) is a kind of RNA binding proteins that plays important roles in many physiological processes. The CIRP has been widely studied in mammals and amphibians since it was first cloned from mammals. On the contrary, there are little reports in teleosts. In this study, the Po CIRP gene of the Japanese flounder was cloned and sequenced. The genomic sequence consists of seven exons and six introns. The putative Po CIRP protein of flounder was 198 amino acid residues long containing the RNA recognition motif(RRM). Phylogenetic analysis showed that the flounder Po CIRP is highly conserved with other teleost CIRPs. The 5' flanking sequence was cloned by genome walking and many transcription factor binding sites were identified. There is a Cp Gs region located in promoter and exon I region and the methylation state is low. Quantitative real-time PCR analysis uncovered that Po CIRP gene was widely expressed in adult tissues with the highest expression level in the ovary. The m RNA of the Po CIRP was maternally deposited and the expression level of the gene was regulated up during the gastrula and neurula stages. In order to gain the information how the protein interacts with m RNA, we performed the modeling of the 3D structure of the flounder Po CIRP. The results showed a cleft existing the surface of the molecular. Taken together, the results indicate that the CIRP is a multifunctional molecular in teleosts and the findings about the structure provide valuable information for understanding the basis of this protein's function.
