| 水稻密码子优化的cry1Ca 基因在大肠杆菌中表达、纯化及抗体制备 |
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| 引用本文： | 林智敏,陈在杰,胡太蛟,胡昌泉,颜静宛,苏军,王锋.水稻密码子优化的cry1Ca 基因在大肠杆菌中表达、纯化及抗体制备[J].福建农业学报,2011,26(1):24-28. |
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| 作者姓名： | 林智敏 陈在杰 胡太蛟 胡昌泉 颜静宛 苏军 王锋 |
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| 作者单位： | 福建省农业科学院农业遗传工程重点实验室,福建,福州,350003 |
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| 基金项目： | 福建省属公益类科研院所基本科研专项，福建省科技计划重点项目 |
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| 摘 要： | 将水稻偏好密码子优化并人工合成抗虫基因cry1Ca通过限制性内切酶酶切的方法构建原核表达载体pET-28bca,成功表达了带6个组氨酸标签的His-cry1ca融合蛋白,其表达量约占菌体总蛋白的24%,表达的融合蛋白主要以包涵体的形式存在.对包涵体蛋白进行可溶性处理,亲和纯化出带6个组氨酸标签的融合蛋白,将其作为抗原免...
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| 关 键 词： | cry1Ca基因 原核表达 融合蛋白 多克隆抗体 |
Expression and purification of rice codon optimized His-cry1Ca fusion protein in E. coli and preparation of polyclonal antibody against cry1Ca |
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| LIN Zhi-min,CHEN Zai-jie,HU Tai-jiao,HU Chang-quan,YAN Jing-wan,SU Jun,WANG Feng.Expression and purification of rice codon optimized His-cry1Ca fusion protein in E. coli and preparation of polyclonal antibody against cry1Ca[J].Fujian Journal of Agricultural Sciences,2011,26(1):24-28. |
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| Authors: | LIN Zhi-min CHEN Zai-jie HU Tai-jiao HU Chang-quan YAN Jing-wan SU Jun WANG Feng |
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| Institution: | LIN Zhi-min,CHEN Zai-jie,HU Tai-jiao,HU Chang-quan,YAN Jing-wan,SU Jun,WANG Feng(Fujian Provincial Key Laboratory of Genetic Engineering for Agriculture,Fujian Academy of Agricultural Sciences,Fuzhou,Fujian 350003,China) |
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| Abstract: | Rice codon optimized cry1Ca gene was cloned into expression vector pET-28b(+) by means of restricted enzymatic digestion.His-cry1Ca fusion protein was highly expressed by SDS-PAGE analysis accumulating more than 24% of the total bacterial proteins.The main expression production was deposited as inclusion body.His-cry1Ca was then purified with Ni-NTA after treatment of solvents.The target protein was used as the antigen to immunize rabbits.ELISA assay showed that the titer of the prepared polyclonal antibody... |
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| Keywords: | cry1Ca gene prokaryotic expression fusion protein polyclonal antibody |
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