| 太平洋牡蛎(Crassostrea gigas)类α-1,2-岩藻糖基转移酶的密码子优化与原核表达 |
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| 作者姓名： | 姚 琳 江艳华 李风铃 朱文嘉 郭莹莹 姜 薇 翟毓秀 王联珠 |
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| 作者单位： | 1. 农业部水产品质量安全检测与评价重点实验室农业部水产品质量安全风险评估实验室(青岛)中国水产科学研究院黄海水产研究所 青岛 266071;2. 潍坊市产品质量检验所 潍坊261061 |
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| 基金项目： | 国家自然科学基金(31101883)和科技部科技基础性工作专项(2013FY113300)共同资助 |
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| 摘 要： | 牡蛎消化组织内存在的类A型血型组织抗原是其特异性富集诺如病毒的主要原因,FUT2(Fucosyltransferase 2,α-1,2-岩藻糖基转移酶)是A型血型组织抗原合成的关键酶.本研究在前期克隆了太平洋牡蛎(Crassostrea gigas)类FUT2基因cDNA全长的基础上,根据大肠杆菌密码子偏爱性优化并合成了类FUT2基因,插入原核表达载体pRSET A构建pRSET-mof,将其转化大肠杆菌BL21,用异丙基-β-D-硫代半乳糖苷(IPTG)诱导.经SDS-PAGE分析显示,在37℃、IPTG终浓度为0.8 mmol/L的条件下,诱导4h后出现大小约为46 kDa的特异性目的条带.利用His亲和层析柱纯化及超滤管浓缩目的蛋白,得到单一条带,说明纯化效果良好.Western blot分析显示,目的蛋白与抗6×His标签单克隆抗体、抗人FUT2单克隆抗体均能发生特异性反应,表明优化后的太平洋牡蛎类FUT2基因在大肠杆菌系统中成功表达.本研究结果为今后研究太平洋牡蛎类FUT2基因的功能,进一步探索牡蛎特异性富集诺如病毒的分子机理奠定了基础.
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| 关 键 词： | 太平洋牡蛎 类FUT2基因 密码子优化 原核表达 |
| 收稿时间： | 2015/3/20 0:00:00 |
| 修稿时间： | 2015/4/27 0:00:00 |
Codon Optimization and Prokaryotic Expression of Fucosyltransferase 2 Like Protein from the Oyster (Crassostrea gigas) |
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| Authors: | YAO Lin JIANG Yanhu LI Fengling ZHU Wenji GUO Yingying JIANG Wei ZHAI Yuxiu and WANG Lianzhu |
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| Institution: | Key Laboratory of Testing and Evaluation for Aquatic Products Safety and Quality, Ministry of Agriculture;Laboratory of Quality & Safety Risk Assessment for Aquatic Products (Qingdao), Ministry of Agriculture; Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071,Key Laboratory of Testing and Evaluation for Aquatic Products Safety and Quality, Ministry of Agriculture;Laboratory of Quality & Safety Risk Assessment for Aquatic Products (Qingdao), Ministry of Agriculture; Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071,Key Laboratory of Testing and Evaluation for Aquatic Products Safety and Quality, Ministry of Agriculture;Laboratory of Quality & Safety Risk Assessment for Aquatic Products (Qingdao), Ministry of Agriculture; Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071,Key Laboratory of Testing and Evaluation for Aquatic Products Safety and Quality, Ministry of Agriculture;Laboratory of Quality & Safety Risk Assessment for Aquatic Products (Qingdao), Ministry of Agriculture; Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071,Key Laboratory of Testing and Evaluation for Aquatic Products Safety and Quality, Ministry of Agriculture;Laboratory of Quality & Safety Risk Assessment for Aquatic Products (Qingdao), Ministry of Agriculture; Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071,Weifang Product Quality Inspection Institute, Weifang 261061,Key Laboratory of Testing and Evaluation for Aquatic Products Safety and Quality, Ministry of Agriculture;Laboratory of Quality & Safety Risk Assessment for Aquatic Products (Qingdao), Ministry of Agriculture; Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071 and Key Laboratory of Testing and Evaluation for Aquatic Products Safety and Quality, Ministry of Agriculture;Laboratory of Quality & Safety Risk Assessment for Aquatic Products (Qingdao), Ministry of Agriculture; Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071 |
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| Abstract: | |
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| Keywords: | Crassostrea gigas FUT2 like gene Codon optimization Prokaryocyte expression |
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