| 半滑舌鳎两种leptin基因的体外重组表达和生物活性分析 |
| |
| 引用本文： | 张雅星,王滨,柳学周,徐永江,史宝,姜燕,崔爱君,张正荣,孙冉冉.半滑舌鳎两种leptin基因的体外重组表达和生物活性分析[J].水产学报,2019,43(11):2279-2289. |
| |
| 作者姓名： | 张雅星 王滨 柳学周 徐永江 史宝 姜燕 崔爱君 张正荣 孙冉冉 |
| |
| 作者单位： | 中国水产科学研究院黄海水产研究所, 农业农村部海洋渔业可持续发展重点实验室, 青岛市海水鱼类种子工程与生物技术重点实验室, 山东 青岛 266071;青岛海洋科学与技术试点国家实验室, 海洋渔业科学与食物产出过程功能实验室, 山东 青岛 266071;上海海洋大学水产与生命学院, 上海 201306,中国水产科学研究院黄海水产研究所, 农业农村部海洋渔业可持续发展重点实验室, 青岛市海水鱼类种子工程与生物技术重点实验室, 山东 青岛 266071;青岛海洋科学与技术试点国家实验室, 海洋渔业科学与食物产出过程功能实验室, 山东 青岛 266071,中国水产科学研究院黄海水产研究所, 农业农村部海洋渔业可持续发展重点实验室, 青岛市海水鱼类种子工程与生物技术重点实验室, 山东 青岛 266071;青岛海洋科学与技术试点国家实验室, 海洋渔业科学与食物产出过程功能实验室, 山东 青岛 266071,中国水产科学研究院黄海水产研究所, 农业农村部海洋渔业可持续发展重点实验室, 青岛市海水鱼类种子工程与生物技术重点实验室, 山东 青岛 266071;青岛海洋科学与技术试点国家实验室, 海洋渔业科学与食物产出过程功能实验室, 山东 青岛 266071,中国水产科学研究院黄海水产研究所, 农业农村部海洋渔业可持续发展重点实验室, 青岛市海水鱼类种子工程与生物技术重点实验室, 山东 青岛 266071;青岛海洋科学与技术试点国家实验室, 海洋渔业科学与食物产出过程功能实验室, 山东 青岛 266071,中国水产科学研究院黄海水产研究所, 农业农村部海洋渔业可持续发展重点实验室, 青岛市海水鱼类种子工程与生物技术重点实验室, 山东 青岛 266071;青岛海洋科学与技术试点国家实验室, 海洋渔业科学与食物产出过程功能实验室, 山东 青岛 266071,中国水产科学研究院黄海水产研究所, 农业农村部海洋渔业可持续发展重点实验室, 青岛市海水鱼类种子工程与生物技术重点实验室, 山东 青岛 266071;青岛海洋科学与技术试点国家实验室, 海洋渔业科学与食物产出过程功能实验室, 山东 青岛 266071;上海海洋大学水产与生命学院, 上海 201306,中国水产科学研究院黄海水产研究所, 农业农村部海洋渔业可持续发展重点实验室, 青岛市海水鱼类种子工程与生物技术重点实验室, 山东 青岛 266071;青岛海洋科学与技术试点国家实验室, 海洋渔业科学与食物产出过程功能实验室, 山东 青岛 266071;上海海洋大学水产与生命学院, 上海 201306,中国水产科学研究院黄海水产研究所, 农业农村部海洋渔业可持续发展重点实验室, 青岛市海水鱼类种子工程与生物技术重点实验室, 山东 青岛 266071;青岛海洋科学与技术试点国家实验室, 海洋渔业科学与食物产出过程功能实验室, 山东 青岛 266071 |
| |
| 基金项目： | 山东省重点研发计划（2018GHY115044）;山东省支持青岛海洋科学与技术试点国家实验室重大科技专项（2018SDKJ0303-1,2018SDKJ0501-2）;中国水产科学研究院基本科研业务费专项（2019GH15）;海水鱼产业技术体系专项（CARS-47）;农业农村部“一带一路”沿线热带国家水产养殖科技合作项目 |
| |
| 摘 要： | 根据半滑舌鳎lepa和lepb编码氨基酸序列合成开放阅读框ORF肽段。利用原核表达载体pEQ30成功构建了半滑舌鳎lepa/pQE30和lepb/pQE30重组质粒,分别转化至大肠杆菌M15后,经IPTG诱导获得了N端含6个组氨酸的半滑舌鳎LepA和LepB重组蛋白。获得的重组蛋白大小均为16 ku,37°C下用0.5 mmol/L的IPTG诱导4 h后目的蛋白表达量最高,主要以包涵体形式存在。检测半滑舌鳎LepA和LepB重组蛋白浓度分别为0.3和0.25 mg/mL。Western blot免疫印迹和质谱分析表明,获得的LepA和LepB重组蛋白均具有免疫活性且序列正确。Ni2+-NTA亲和层析柱纯化可获得高纯度的半滑舌鳎LepA和LepB重组蛋白。离体孵育实验表明,获得的半滑舌鳎LepA和LepB重组蛋白能显著抑制半滑舌鳎下丘脑lepa、lepb和gnrh3 mRNA的表达水平,表明获得的重组蛋白具有明显的生物活性。研究结果可为探究leptin在半滑舌鳎生长发育中的调控机制提供重要参考。
|
| 关 键 词： | 半滑舌鳎 leptin 原核表达 生物活性 |
| 收稿时间： | 2018/12/27 0:00:00 |
| 修稿时间： | 2019/5/28 0:00:00 |
Recombinant expression and bioactivity analysis of two leptin genes of Cynoglossus semilaevis |
| |
| ZHANG Yaxing,WANG Bin,LIU Xuezhou,XU Yongjiang,SHI Bao,JIANG Yan,CUI Aijun,ZHANG Zhengrong and SUN Ranran.Recombinant expression and bioactivity analysis of two leptin genes of Cynoglossus semilaevis[J].Journal of Fisheries of China,2019,43(11):2279-2289. |
| |
| Authors: | ZHANG Yaxing WANG Bin LIU Xuezhou XU Yongjiang SHI Bao JIANG Yan CUI Aijun ZHANG Zhengrong and SUN Ranran |
| |
| Institution: | Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture and Rural Affairs, Qingdao Key Laboratory for Marine Fish Breeding and Biotechnology, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;Laboratory for Marine Fisheries Science and Food Production Processes, Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266071, China;College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China,Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture and Rural Affairs, Qingdao Key Laboratory for Marine Fish Breeding and Biotechnology, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;Laboratory for Marine Fisheries Science and Food Production Processes, Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266071, China,Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture and Rural Affairs, Qingdao Key Laboratory for Marine Fish Breeding and Biotechnology, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;Laboratory for Marine Fisheries Science and Food Production Processes, Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266071, China,Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture and Rural Affairs, Qingdao Key Laboratory for Marine Fish Breeding and Biotechnology, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;Laboratory for Marine Fisheries Science and Food Production Processes, Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266071, China,Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture and Rural Affairs, Qingdao Key Laboratory for Marine Fish Breeding and Biotechnology, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;Laboratory for Marine Fisheries Science and Food Production Processes, Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266071, China,Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture and Rural Affairs, Qingdao Key Laboratory for Marine Fish Breeding and Biotechnology, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;Laboratory for Marine Fisheries Science and Food Production Processes, Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266071, China,Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture and Rural Affairs, Qingdao Key Laboratory for Marine Fish Breeding and Biotechnology, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;Laboratory for Marine Fisheries Science and Food Production Processes, Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266071, China;College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China,Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture and Rural Affairs, Qingdao Key Laboratory for Marine Fish Breeding and Biotechnology, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;Laboratory for Marine Fisheries Science and Food Production Processes, Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266071, China;College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China and Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture and Rural Affairs, Qingdao Key Laboratory for Marine Fish Breeding and Biotechnology, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;Laboratory for Marine Fisheries Science and Food Production Processes, Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266071, China |
| |
| Abstract: | |
| |
| Keywords: | Cynoglossus semilaevis leptin prokaryotic expression bioactivity analysis |
| 本文献已被 CNKI 等数据库收录！ |
| 点击此处可从《水产学报》浏览原始摘要信息 |
| 点击此处可从《水产学报》下载免费的PDF全文 |
|
