| 苏云金芽胞杆菌cry1Ie新基因的克隆、表达与生物活性测定 |
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| 引用本文： | 余宗兰,贺利业,龚莉,黄刚辉,李平,郑爱萍.苏云金芽胞杆菌cry1Ie新基因的克隆、表达与生物活性测定[J].植物保护学报,2016,43(2):201-206. |
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| 作者姓名： | 余宗兰 贺利业 龚莉 黄刚辉 李平 郑爱萍 |
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| 作者单位： | 四川农业大学水稻研究所,温江611130;四川农业大学,西南作物基因资源与遗传改良教育部重点实验室,雅安625014 |
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| 基金项目： | 国家重大转基因专项(2011ZX08009-003-001-009) |
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| 摘 要： | 为获得新的鳞翅目害虫杀虫基因,根据已知cry1I类基因编码区设计简并引物,采用直接克隆法,以苏云金芽胞杆菌(Bacillus thuringiensis,Bt)菌株BN23-5质粒DNA为模板进行扩增,并对得到的基因进行鉴定和分析。结果表明:克隆得到一个完整的cry1Ie基因,全长2 160 bp,由719个氨基酸组成,该氨基酸序列与已知的4种Cry1Ie蛋白不同,与Cry1Ie2和Cry1Ie3的氨基酸序列同源性最高,为95.4%,被国际Bt杀虫晶体蛋白基因命名委员会命名为Cry1Ie5(登录号为KJ710646)。将该基因插入表达载体p ET-28a,转化大肠杆菌BL21(DE3),IPTG低温诱导成功表达,SDS-PAGE电泳验证其大小为81 k D,与预测的分子量相符合。生物活性测定表明,Cry1Ie5表达的包涵体蛋白对小菜蛾和亚洲玉米螟具有杀虫活性,LC_(50)分别为0.43μg/m L和48.39μg/m L;对棉铃虫的致死率不高,但能明显抑制其生长;对甜菜夜蛾没有杀虫活性。
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| 关 键 词： | 苏云金芽胞杆菌 cry1 Ie5基因 鳞翅目害虫 克隆 |
| 收稿时间： | 2014/10/10 0:00:00 |
Cloning, expression, and biological activity of a new cry1Ie gene from Bacillus thuringiensis strain |
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| Yu Zonglan,He Liye,Gong Li,Huang Ganghui,Li Ping and Zheng Aiping.Cloning, expression, and biological activity of a new cry1Ie gene from Bacillus thuringiensis strain[J].Acta Phytophylacica Sinica,2016,43(2):201-206. |
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| Authors: | Yu Zonglan He Liye Gong Li Huang Ganghui Li Ping and Zheng Aiping |
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| Institution: | Rice Research Institute, Sichuan Agricultural University, Wenjiang 611130, Sichuan Province, China;Key Laboratory of Southwest Crop Gene Resource & Genetic Improvement of Ministry of Education, Sichuan Agricultural University, Ya'an 625014, Sichuan Province, China,Rice Research Institute, Sichuan Agricultural University, Wenjiang 611130, Sichuan Province, China;Key Laboratory of Southwest Crop Gene Resource & Genetic Improvement of Ministry of Education, Sichuan Agricultural University, Ya'an 625014, Sichuan Province, China,Rice Research Institute, Sichuan Agricultural University, Wenjiang 611130, Sichuan Province, China;Key Laboratory of Southwest Crop Gene Resource & Genetic Improvement of Ministry of Education, Sichuan Agricultural University, Ya'an 625014, Sichuan Province, China,Rice Research Institute, Sichuan Agricultural University, Wenjiang 611130, Sichuan Province, China;Key Laboratory of Southwest Crop Gene Resource & Genetic Improvement of Ministry of Education, Sichuan Agricultural University, Ya'an 625014, Sichuan Province, China,Rice Research Institute, Sichuan Agricultural University, Wenjiang 611130, Sichuan Province, China;Key Laboratory of Southwest Crop Gene Resource & Genetic Improvement of Ministry of Education, Sichuan Agricultural University, Ya'an 625014, Sichuan Province, China and Rice Research Institute, Sichuan Agricultural University, Wenjiang 611130, Sichuan Province, China;Key Laboratory of Southwest Crop Gene Resource & Genetic Improvement of Ministry of Education, Sichuan Agricultural University, Ya'an 625014, Sichuan Province, China |
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| Abstract: | |
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| Keywords: | Bacillus thuringiensis cry1Ie5 Lepidoptera pest clone |
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