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半滑舌鳎(Cynoglossus semilaevis)Patched 1基因的克隆和表达分析
引用本文:董忠典,祁茜,张宁,邵长伟,张丽燕,温海深,陈松林.半滑舌鳎(Cynoglossus semilaevis)Patched 1基因的克隆和表达分析[J].海洋学报(英文版),2016,35(6):19-28.
作者姓名:董忠典  祁茜  张宁  邵长伟  张丽燕  温海深  陈松林
作者单位:中国海洋大学, 中国青岛 266003;中国水产科学研究院 黄海水产研究所/农业部海洋渔业可持续发展重点实验室, 中国青岛 266071;青岛海洋科学与技术国家实验室/海洋渔业科学与食物产出过程功能实验室, 中国青岛 266237,中国海洋大学, 中国青岛 266003;中国水产科学研究院 黄海水产研究所/农业部海洋渔业可持续发展重点实验室, 中国青岛 266071;青岛海洋科学与技术国家实验室/海洋渔业科学与食物产出过程功能实验室, 中国青岛 266237,中国海洋大学, 中国青岛 266003;中国水产科学研究院 黄海水产研究所/农业部海洋渔业可持续发展重点实验室, 中国青岛 266071;青岛海洋科学与技术国家实验室/海洋渔业科学与食物产出过程功能实验室, 中国青岛 266237,中国水产科学研究院 黄海水产研究所/农业部海洋渔业可持续发展重点实验室, 中国青岛 266071;青岛海洋科学与技术国家实验室/海洋渔业科学与食物产出过程功能实验室, 中国青岛 266237,中国水产科学研究院 黄海水产研究所/农业部海洋渔业可持续发展重点实验室, 中国青岛 266071;青岛海洋科学与技术国家实验室/海洋渔业科学与食物产出过程功能实验室, 中国青岛 266237,中国海洋大学, 中国青岛 266003,中国水产科学研究院 黄海水产研究所/农业部海洋渔业可持续发展重点实验室, 中国青岛 266071;青岛海洋科学与技术国家实验室/海洋渔业科学与食物产出过程功能实验室, 中国青岛 266237
摘    要:Hedgehog(Hh)信号通路在脊椎动物和非脊椎动物发育过程中细胞信号传导方面发挥重要的作用。Patched 1 (Ptc1)基因是Hh的受体,通过和Hh配体结合调节Hh信号通路。本研究克隆和鉴定了半滑舌鳎的Ptc1基因(csptc1),该基因cDNA全长为5212 nt,编码蛋白包括1543个氨基酸残基,序列比对和进化分析显示该蛋白在进化过程中较为保守。荧光定量显示该基因在脑,肝脏,心脏,鳃,肠,脾脏、肾脏和性腺组织中普遍表达,在精巢中的表达水平显著高于卵巢。精巢中的原位杂交信号主要位于该基因主要在初级精母细胞,次级精母细胞和支持细胞中,而在卵母细胞中的信号较弱。Csptc1基因在卵巢中的甲基化水平高于在雄鱼和伪雄鱼精巢中。这一研究结果可能说明csptc1基因参与了 Desert Hedgehog (DHH)基因维持雄鱼和伪雄鱼的生殖细胞系以及精子发生等生物学过程。

关 键 词:半滑舌鳎  Patched  1  性别  快速扩增cDNA末端  实时荧光定量PCR  甲基化
收稿时间:2015/6/11 0:00:00
修稿时间:2015/12/3 0:00:00

Molecular characterization and expression analysis of Patched 1 gene in the half-smooth tongue sole (Cynoglossus semilaevis)
DONG Zhongdian,QI Qian,ZHANG Ning,SHAO Changwei,ZHANG Liyan,WEN Haishen and CHEN Songlin.Molecular characterization and expression analysis of Patched 1 gene in the half-smooth tongue sole (Cynoglossus semilaevis)[J].Acta Oceanologica Sinica,2016,35(6):19-28.
Authors:DONG Zhongdian  QI Qian  ZHANG Ning  SHAO Changwei  ZHANG Liyan  WEN Haishen and CHEN Songlin
Institution:Fisheries College, Ocean University of China, Qingdao 266003, China;Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;Function Laboratory for Marine Fisheries Science and Food Production Processes, National Laboratory for Marine Science and Technology, Qingdao 266237, China,Fisheries College, Ocean University of China, Qingdao 266003, China;Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;Function Laboratory for Marine Fisheries Science and Food Production Processes, National Laboratory for Marine Science and Technology, Qingdao 266237, China,Fisheries College, Ocean University of China, Qingdao 266003, China;Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;Function Laboratory for Marine Fisheries Science and Food Production Processes, National Laboratory for Marine Science and Technology, Qingdao 266237, China,Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;Function Laboratory for Marine Fisheries Science and Food Production Processes, National Laboratory for Marine Science and Technology, Qingdao 266237, China,Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;Function Laboratory for Marine Fisheries Science and Food Production Processes, National Laboratory for Marine Science and Technology, Qingdao 266237, China,Fisheries College, Ocean University of China, Qingdao 266003, China and Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;Function Laboratory for Marine Fisheries Science and Food Production Processes, National Laboratory for Marine Science and Technology, Qingdao 266237, China
Abstract:The signaling molecule hedgehog (Hh) is essential for cellular signaling required for body pattern formation in embryonic and individual development in vertebrate and invertebrates. Patched 1 (Ptc1), a receptor of Hh, mediates the activity of Hh by binging the Hh ligands. In this study, the Ptc1 of half-smooth tongue sole Cynoglossus semilaevis (CsPtc1) was cloned and characterized. The full-length of CsPtc1 cDNA is 5 212 nucleotides (nt), and encodes a protein with 1 543 amino acid residues. CsPtc1 shared many features with Ptc1 of other telesot species. Real-time quantitative PCR revealed CsPtc1 mRNA expressed in brain, liver, heart, gill, intestines, spleen, gonad and kidney, with expression level in testis significantly higher than in ovary. In testis, hybridization signals were mainly detected in primary spermatocytes, secondary spermatocytes, and sertoli cells, while weak signals were found in oocytes. Correspondingly, the degree of methylation is higher in female than in male and pseudo-male. Results indicate that CsPtc1 may be involved in Desert Hedgehog (DHH) maintenance of the male and pseudo-male germ line and spermatogenesis.
Keywords:half-smooth tongue sole  male and female  Patched 1  RACE  qRT-PCR  methylation
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