在糙皮侧耳(Pleurotus ostreatus)的全基因组中鉴定出14个高速泳动蛋白(high-mobility group box,HMG-box)的编码基因(PoHMG1~PoHMG14),对其进行特征、结构与系统进化分析,发现所有的HMGbox转录因子蛋白的三级结构高度相似,相对分子质量...在糙皮侧耳(Pleurotus ostreatus)的全基因组中鉴定出14个高速泳动蛋白(high-mobility group box,HMG-box)的编码基因(PoHMG1~PoHMG14),对其进行特征、结构与系统进化分析,发现所有的HMGbox转录因子蛋白的三级结构高度相似,相对分子质量为6893.79~60397.29,蛋白PoHMG6、PoHMG11、PoHMG12与PoHMG13之间的亲缘关系较近。基于糙皮侧耳在菌丝、原基、幼嫩子实体和成熟子实体时期的转录组数据,对筛选到的转录因子基因PoHMG11进行克隆分析并探究其功能及表达特性,发现该基因全长序列1517 bp,编码489个氨基酸,含有一个HMG-box结构域。通过大肠杆菌原核表达载体体外诱导表达出相对分子质量为5.5×104的蛋白,该重组蛋白在1 mol·L^(-1)IPTG、25℃诱导表达6 h时表达量最高。利用实时荧光定量PCR(qRT-PCR)分析糙皮侧耳在菌丝、原基、幼嫩子实体和成熟子实体阶段PoHMG11基因的相对表达量,发现该基因在成熟糙皮侧耳子实体中的表达量比其他生长发育时期显著升高5倍以上,推测该基因可能参与调控糙皮侧耳子实体成熟过程。展开更多
为探明Sox基因在物种进化中的保守性及其性别分型,以大鲵雌、雄个体为材料,参照Sox基因HMG-box保守区的序列,设计简并引物扩增,两因子均在雌、雄个体内得到217 bp PCR扩增产物,不存在性别差异。将二者编码的氨基酸序列与NCBI中其他物种...为探明Sox基因在物种进化中的保守性及其性别分型,以大鲵雌、雄个体为材料,参照Sox基因HMG-box保守区的序列,设计简并引物扩增,两因子均在雌、雄个体内得到217 bp PCR扩增产物,不存在性别差异。将二者编码的氨基酸序列与NCBI中其他物种Sox基因编码的氨基酸序列进行比对,其中一个基因与人、斑马鱼、家鼠、鸡、热带爪蟾、扬子鳄Sox11基因的同源性均为90%,另一个基因与罗非鱼、鸭嘴兽、家鼠、红鳍东方鲀、斑马鱼、鸡、人、非洲爪蟾和大鲵Sox14基因的同源性均为90%,故根据大鲵的学名,将这2个基因分别命名为adSox11和adSox14c。adSox11和adSox14c在大鲵精巢、卵巢、肾脏和心脏中均有不同程度表达,而在胃和肝脏中几乎不表达。展开更多
Background SRY-related HMG-box 17 （SOX17） encodes a member of the SOX （SRY-related HMG-box） family of transcription factors involved in the regulation of embryonic development and in the determination of the cell ...Background SRY-related HMG-box 17 （SOX17） encodes a member of the SOX （SRY-related HMG-box） family of transcription factors involved in the regulation of embryonic development and in the determination of the cell fate. Recently, it was considered as a tumor suppressor gene to inhibit canonical Wnt/β-catenin signaling pathway in several malignancies. However, the function of SOX17 in thyroid cancer was unknown. Therefore, we investigated the epigenetic changes and the function of SOX17 in thyroid cancer. Methods The methylation status of the promoter region of SOX17 was detected using methylation-specific PCR in 63 papillary thyroid carcinoma （PTC） tissue, 10 normal thyroid tissue, and two thyroid cancer cell lines. Semi-quantitative RT-PCR was used to assess mRNA expression of SOX17 before and after 5-aza-2＇-deoxycytidine treatment in thyroid cancer cell lines. Expression of SOX17 and β-catenin were detected by immunohistochemistry in PTC and adjacent tissue. Luciferase reporter assay, colony formation, transfection, and Western blotting were employed to analyze the effect of SOX17 on thyroid cancer cell proliferation and the function of SOX17 in the Wnt signal pathway. Results Loss of SOX17 expression was correlated to the promoter region hypermethylation in thyroid cancer cell lines. Re-expression of SOX17 was found in TPC-1 cell line after 5-aza-2＇-deoxycytidine treatment. In primary thyroid cancer, 60.3% （38/63） were methylated and 39.7% （25/63） unmethylated. But no methylation was found in noncancerous thyroid tissues. Methylation of SOX17 was associated reversely with β-catenin expression in the cytoplasm or nucleus significantly in the PTC （P 〈0.05）. Colony formation was inhibited by re-expression of SOX17 in TPC-1 cells. SOX17 suppressed the Wnt signaling pathway and the HMG domain was essential for this effect. Conclusions SOX17 was frequently methylated in human PTC. Loss of SOX17 expression was induced by promoter region hypermethylation. SOX17 inhibited thyroid cancer proliferation. Methylation of SOX17 activated the Wnt signaling pathway in human thyroid cancer.展开更多
We amplified the 197 bp HMG box sequence by using Polymerase Chain Reaction (PCR) with primers according to the known SRY sequence from male raccoon genomic DNA, and then cloned and sequenced. The nucleic acid and ami...We amplified the 197 bp HMG box sequence by using Polymerase Chain Reaction (PCR) with primers according to the known SRY sequence from male raccoon genomic DNA, and then cloned and sequenced. The nucleic acid and amino acid sequence comparison between raccoon and other mammals revealed high conservation of the SRY HMG box in mammals (about 80%), which implied that the DNA binding activity was crucial to mediate the action of SRY in sex determination. The variation of raccoon HMG box sequence mainly occurred on purine by replacement and missense mutation, which implicated that only the HMG box protein with advanced structure had efficient activity. The homology and difference between mammals HMG box was accorded to the evolution systematic tree.展开更多
Sox(Sry-related high mobility group box)基因是由众多具有HMG-box保守基序构成的超基因家族,是与位于雄性动物Y染色体上的Sry基因同源的家族基因,在很多动物中都有表达。由于其编码的蛋白质具有结合DNA的能力,因而认为Sox基因家族是...Sox(Sry-related high mobility group box)基因是由众多具有HMG-box保守基序构成的超基因家族,是与位于雄性动物Y染色体上的Sry基因同源的家族基因,在很多动物中都有表达。由于其编码的蛋白质具有结合DNA的能力,因而认为Sox基因家族是一类重要的转录调控因子。在个体发育过程中,Sox基因广泛参与了动物的早期胚胎发育、性别决定和分化、神经系统发育、软骨及多种组织器官的形成,具有重要的生物学功能。主要对Sox家族基因的功能及其研究进展进行简要的综述。展开更多
Four different Sox genes in Amur sturgeon Acipenser schrenckii and five Sox genes in chum salmon Oncorhynchus keta were identified for the first time in this paper, and there was no sexual difference for these genes i...Four different Sox genes in Amur sturgeon Acipenser schrenckii and five Sox genes in chum salmon Oncorhynchus keta were identified for the first time in this paper, and there was no sexual difference for these genes in these two fishes. For Amur sturgeon, the encoded amino acid sequences exhibited 98%, 98%, 98%, 86% of sequence homology with those reported for human SOX HMG box, respectively, and for chum salmon, the encoded amino acid sequences exhibited 93%,98%,98%,98%,96% of sequence homology with those reported for human SOX HMG box, respectively. Two different forms of Sox9 gene in chum salmon were found, and Sox9 contained a 303 bp and 258 bp introns in the HMG box for chum salmon respectively while 572 bp for Amur sturgeon. No clear similarity was observed between the introns in the Sox9 HMG box of different fishes, but the introns occur in the HMG box region between the second and third bases of the codon for arginine at residue 42 in HMG box for the three fishes, and obey the rule of “GT AG”. and the encoded amino acid sequences in the three Sox9 genes HMG box are very conservative in the two fishes (100% identification). Identification of these genes is a potential step in understanding development regulations including sex determination in Amur sturgeon.展开更多
文摘为探明Sox基因在物种进化中的保守性及其性别分型,以大鲵雌、雄个体为材料,参照Sox基因HMG-box保守区的序列,设计简并引物扩增,两因子均在雌、雄个体内得到217 bp PCR扩增产物,不存在性别差异。将二者编码的氨基酸序列与NCBI中其他物种Sox基因编码的氨基酸序列进行比对,其中一个基因与人、斑马鱼、家鼠、鸡、热带爪蟾、扬子鳄Sox11基因的同源性均为90%,另一个基因与罗非鱼、鸭嘴兽、家鼠、红鳍东方鲀、斑马鱼、鸡、人、非洲爪蟾和大鲵Sox14基因的同源性均为90%,故根据大鲵的学名,将这2个基因分别命名为adSox11和adSox14c。adSox11和adSox14c在大鲵精巢、卵巢、肾脏和心脏中均有不同程度表达,而在胃和肝脏中几乎不表达。
文摘Background SRY-related HMG-box 17 （SOX17） encodes a member of the SOX （SRY-related HMG-box） family of transcription factors involved in the regulation of embryonic development and in the determination of the cell fate. Recently, it was considered as a tumor suppressor gene to inhibit canonical Wnt/β-catenin signaling pathway in several malignancies. However, the function of SOX17 in thyroid cancer was unknown. Therefore, we investigated the epigenetic changes and the function of SOX17 in thyroid cancer. Methods The methylation status of the promoter region of SOX17 was detected using methylation-specific PCR in 63 papillary thyroid carcinoma （PTC） tissue, 10 normal thyroid tissue, and two thyroid cancer cell lines. Semi-quantitative RT-PCR was used to assess mRNA expression of SOX17 before and after 5-aza-2＇-deoxycytidine treatment in thyroid cancer cell lines. Expression of SOX17 and β-catenin were detected by immunohistochemistry in PTC and adjacent tissue. Luciferase reporter assay, colony formation, transfection, and Western blotting were employed to analyze the effect of SOX17 on thyroid cancer cell proliferation and the function of SOX17 in the Wnt signal pathway. Results Loss of SOX17 expression was correlated to the promoter region hypermethylation in thyroid cancer cell lines. Re-expression of SOX17 was found in TPC-1 cell line after 5-aza-2＇-deoxycytidine treatment. In primary thyroid cancer, 60.3% （38/63） were methylated and 39.7% （25/63） unmethylated. But no methylation was found in noncancerous thyroid tissues. Methylation of SOX17 was associated reversely with β-catenin expression in the cytoplasm or nucleus significantly in the PTC （P 〈0.05）. Colony formation was inhibited by re-expression of SOX17 in TPC-1 cells. SOX17 suppressed the Wnt signaling pathway and the HMG domain was essential for this effect. Conclusions SOX17 was frequently methylated in human PTC. Loss of SOX17 expression was induced by promoter region hypermethylation. SOX17 inhibited thyroid cancer proliferation. Methylation of SOX17 activated the Wnt signaling pathway in human thyroid cancer.
文摘We amplified the 197 bp HMG box sequence by using Polymerase Chain Reaction (PCR) with primers according to the known SRY sequence from male raccoon genomic DNA, and then cloned and sequenced. The nucleic acid and amino acid sequence comparison between raccoon and other mammals revealed high conservation of the SRY HMG box in mammals (about 80%), which implied that the DNA binding activity was crucial to mediate the action of SRY in sex determination. The variation of raccoon HMG box sequence mainly occurred on purine by replacement and missense mutation, which implicated that only the HMG box protein with advanced structure had efficient activity. The homology and difference between mammals HMG box was accorded to the evolution systematic tree.
文摘Sox(Sry-related high mobility group box)基因是由众多具有HMG-box保守基序构成的超基因家族,是与位于雄性动物Y染色体上的Sry基因同源的家族基因,在很多动物中都有表达。由于其编码的蛋白质具有结合DNA的能力,因而认为Sox基因家族是一类重要的转录调控因子。在个体发育过程中,Sox基因广泛参与了动物的早期胚胎发育、性别决定和分化、神经系统发育、软骨及多种组织器官的形成,具有重要的生物学功能。主要对Sox家族基因的功能及其研究进展进行简要的综述。
文摘Four different Sox genes in Amur sturgeon Acipenser schrenckii and five Sox genes in chum salmon Oncorhynchus keta were identified for the first time in this paper, and there was no sexual difference for these genes in these two fishes. For Amur sturgeon, the encoded amino acid sequences exhibited 98%, 98%, 98%, 86% of sequence homology with those reported for human SOX HMG box, respectively, and for chum salmon, the encoded amino acid sequences exhibited 93%,98%,98%,98%,96% of sequence homology with those reported for human SOX HMG box, respectively. Two different forms of Sox9 gene in chum salmon were found, and Sox9 contained a 303 bp and 258 bp introns in the HMG box for chum salmon respectively while 572 bp for Amur sturgeon. No clear similarity was observed between the introns in the Sox9 HMG box of different fishes, but the introns occur in the HMG box region between the second and third bases of the codon for arginine at residue 42 in HMG box for the three fishes, and obey the rule of “GT AG”. and the encoded amino acid sequences in the three Sox9 genes HMG box are very conservative in the two fishes (100% identification). Identification of these genes is a potential step in understanding development regulations including sex determination in Amur sturgeon.
