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中国明对虾calcineurin B基因的克隆及其在竞争行为中作用的初探
引用本文:孙犁,卢霞,孔杰,孟宪红,闫允君,李波波,仲伟鹏,陈宝龙,曹宝祥,罗坤,栾生.中国明对虾calcineurin B基因的克隆及其在竞争行为中作用的初探[J].水产学报,2020,44(1):11-20.
作者姓名:孙犁  卢霞  孔杰  孟宪红  闫允君  李波波  仲伟鹏  陈宝龙  曹宝祥  罗坤  栾生
作者单位: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,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,山东青岛 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,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,山东青岛 266071
基金项目:国家自然科学基金(31572616);泰山学者种业人才团队项目;现代农业产业技术体系专项(CARS-48);国家自然科学基金联合基金(U1706203);国家重点研发计划(2018YFD0901301)
摘    要:中国明对虾在养殖中表现出较强的竞争行为,对其生长性能产生了较大的影响,然而,到目前为止其发生的分子机制尚不清楚。钙调神经磷酸酶(calcineurin,CN)是高度保守的Ca2+/钙调蛋白(calmodulin,CaM)依赖性丝氨酸/苏氨酸磷酸酶,由催化亚基(CNA)和调节亚基(CN-B)组成,是参与许多重要生理过程的多功能蛋白质。CNB在Ca2+/CaM的介导下主要在动物的中枢神经系统发挥重要作用。另外,前期通过比较转录组分析筛选出的与中国明对虾竞争行为相关的候选基因中包括CNB基因。为了进一步明确CNB在中国明对虾竞争过程中的作用,实验通过RACE技术克隆了中国明对虾CNB基因(FcCN-B)的全长cDNA序列,并利用Real-time PCR技术分析了其在高竞争能力组(HCG)和低竞争能力组(LCG)组间不同组织(神经节、心脏、胃、肝胰腺和肠)中的表达情况。结果显示,FcCN-B的cDNA全长序列为2867 bp,包括95 bp的5′非编码区(UTR),540 bp的开放阅读框(ORF),和2232 bp的3′UTR,其中ORF中具有4个保守的EF-hand Ca2+结合结构域。蛋白质同源性分析显示,FcCN-B的氨基酸序列与其他物种具有较高的同源性(78.8%~93.8%),其中最高的是中华绒螯蟹(93.8%)和黑腹果蝇(90.5%);系统进化关系分析显示,脊椎动物和无脊椎动物分别独立为一支,且中国明对虾与中华绒螯蟹单独聚为一支,之后与黑腹果蝇聚类关系最近,提示FcCN-B在中国明对虾中可能具有与其在中华绒螯蟹和果蝇中相类似的功能。Real-time PCR定量结果显示,FcCN-B在HCG组的神经节中的表达极显著高于LCG组,而其在HCG组的心脏中的表达极显著低于LCG组。研究结果表明,calcineurin B基因在中国明对虾的竞争行为中可能发挥一定的作用,将为解析中国明对虾竞争行为的分子机制奠定重要的基础。

关 键 词:中国明对虾  竞争行为  钙调神经磷酸酶  基因克隆  表达分析
收稿时间:2019/1/11 0:00:00
修稿时间:2019/4/10 0:00:00

Identification and characterization of the calcineurin B in Fenneropenaeus chinensis: its implications in competitive ability
SUN Li,LU Xi,KONG Jie,MENG Xianhong,YAN Yunjun,LI Bobo,ZHONG Weipeng,CHEN Baolong,CAO Baoxiang,LUO Kun and LUAN Sheng.Identification and characterization of the calcineurin B in Fenneropenaeus chinensis: its implications in competitive ability[J].Journal of Fisheries of China,2020,44(1):11-20.
Authors:SUN Li  LU Xi  KONG Jie  MENG Xianhong  YAN Yunjun  LI Bobo  ZHONG Weipeng  CHEN Baolong  CAO Baoxiang  LUO Kun and LUAN Sheng
Institution:College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China;Key Laboratory for Sustainable Utilization of Marine Fisheries Resources, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China,Key Laboratory for Sustainable Utilization of Marine Fisheries Resources, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266071, China,Key Laboratory for Sustainable Utilization of Marine Fisheries Resources, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266071, China,Key Laboratory for Sustainable Utilization of Marine Fisheries Resources, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266071, China,Key Laboratory for Sustainable Utilization of Marine Fisheries Resources, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266071, China,College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China;Key Laboratory for Sustainable Utilization of Marine Fisheries Resources, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China,Key Laboratory for Sustainable Utilization of Marine Fisheries Resources, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China,Key Laboratory for Sustainable Utilization of Marine Fisheries Resources, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266071, China,Key Laboratory for Sustainable Utilization of Marine Fisheries Resources, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266071, China,Key Laboratory for Sustainable Utilization of Marine Fisheries Resources, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266071, China and Key Laboratory for Sustainable Utilization of Marine Fisheries Resources, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266071, China
Abstract:
Keywords:Fenneropenaeus chinensis  competitive behavior  calcineurin B  gene cloning  expression analysis
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