| 人工改造基因Cry1Abm的合成、原核表达及其抗虫鉴定 |
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| 引用本文： | 岳润清,铁双贵,陈小洁,齐建双,韩小花,燕树峰,徐玉格,林鸿.人工改造基因Cry1Abm的合成、原核表达及其抗虫鉴定[J].玉米科学,2013,21(6):21-25,30. |
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| 作者姓名： | 岳润清 铁双贵 陈小洁 齐建双 韩小花 燕树峰 徐玉格 林鸿 |
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| 作者单位： | 河南省农业科学院粮食作物研究所/河南省玉米生物学重点实验室, 郑州 450002;河南省农业科学院粮食作物研究所/河南省玉米生物学重点实验室, 郑州 450002;河南师范大学生命科学学院, 河南 新乡 453007;河南省农业科学院粮食作物研究所/河南省玉米生物学重点实验室, 郑州 450002;河南省农业科学院粮食作物研究所/河南省玉米生物学重点实验室, 郑州 450002;河南省农业科学院粮食作物研究所/河南省玉米生物学重点实验室, 郑州 450002;河南师范大学生命科学学院, 河南 新乡 453007;郑州大学生物工程系, 郑州 450001 |
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| 基金项目： | 国家转基因新品种培育重大专项（2011ZX08003-001） |
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| 摘 要： | 根据植物密码子的偏好性及使用频率，对苏云金芽孢杆菌Cry1Ab野生型基因的编码区序列进行优化和改造，改造后的Cry1Abm基因序列与原始序列同源性为66.2%，G+C含量由37.3%提高到62.7%。人工合成Cry1Abm基因，并将人工合成的改造后的Cry1Abm基因构建到原核表达载体pET28b中，构建原核表达载体pETAbm。将原核表达载体转入大肠杆菌BL21（DE3）中进行诱导表达，用诱导表达的蛋白进行饲虫（玉米螟）实验。结果表明，该蛋白对幼虫具有很强毒性，幼虫的死亡率高达86.63%，同时，存活幼虫的生长发育也受到明显抑制。该基因可以作为杀虫工程及培育转基因抗虫作物的候选基因。
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| 关 键 词： | Cry1Abm 人工合成 原核表达 大肠杆菌 玉米螟 |
| 收稿时间： | 6/6/2013 12:00:00 AM |
Synthesis of the Modified Cry1Abm Gene, Expressionin E. coli and Bioassay |
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| YUE Run-qing,TIE Shuang-gui,CHEN Xiao-jie,QI Jian-shuang,HAN Xiao-hu,YAN Shu-feng,XU Yu-ge and LIN Hong.Synthesis of the Modified Cry1Abm Gene, Expressionin E. coli and Bioassay[J].Journal of Maize Sciences,2013,21(6):21-25,30. |
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| Authors: | YUE Run-qing TIE Shuang-gui CHEN Xiao-jie QI Jian-shuang HAN Xiao-hu YAN Shu-feng XU Yu-ge and LIN Hong |
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| Institution: | Cereal Crops Institute, Henan Academy of Agricultural Sciences/Henan Provincial Key Lab. of Maize Biology, Zhengzhou 450002;Cereal Crops Institute, Henan Academy of Agricultural Sciences/Henan Provincial Key Lab. of Maize Biology, Zhengzhou 450002;College of Life science of Henan Normal University, Xinxiang 453007;Cereal Crops Institute, Henan Academy of Agricultural Sciences/Henan Provincial Key Lab. of Maize Biology, Zhengzhou 450002;Cereal Crops Institute, Henan Academy of Agricultural Sciences/Henan Provincial Key Lab. of Maize Biology, Zhengzhou 450002;Cereal Crops Institute, Henan Academy of Agricultural Sciences/Henan Provincial Key Lab. of Maize Biology, Zhengzhou 450002;College of Life science of Henan Normal University, Xinxiang 453007;Bioengineering Department of Zhengzhou University, Zhengzhou 450001, China |
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| Abstract: | The coding regions of wild Bacillus thuringiensis Cry1Ab was optimized and modified according to plant preferred codons and frequency. The modified Cry1Abm gene had 66.2 percent homology with the native cry1Ab gene, and G+C content was raised from 37.3% to 62.7%. The modified Cry1Abm gene was artificially synthesized. Modified Cry1Abm gene was cloned into prokaryotic expression vector pET28bm, to construct plasmid pETAbm. The protein expression in E. coli BL21(DE3) was confirmed by SDS-PAGE analysis. The results of insect assay with Asian corn borer showed the protein crude expression products of Cry1Abm gene in E. coli had a toxicity to corn borer. The mortality of insect larvae was accounted for 86.63%, and the growth of the survival larvae was seriously inhibited. |
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| Keywords: | cry1Abm Synthesis Prokaryotic expression E coli Asian corn borer |
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