| 苏云金芽孢杆菌Cry8Ca2蛋白的纯化与活性的研究 |
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| 引用本文： | 吴洪福,;张彦彩;郭淑元;张杰;王容燕;高继国.苏云金芽孢杆菌Cry8Ca2蛋白的纯化与活性的研究[J].植物保护,2007,33(4):29-32. |
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| 作者姓名： | 吴洪福 ;张彦彩;郭淑元;张杰;王容燕;高继国 |
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| 作者单位： | 中国农业科学院植物保护研究所植物病虫害生物学国家重点实验室，北京理工大学生命科学与技术学院，北京理工大学生命科学与技术学院，中国农业科学院植物保护研究所植物病虫害生物学国家重点实验室，河北省农林科学院植物保护研究所，东北农业大学生命科学院 北京100094，东北农业大学生命科学院，哈尔滨150030，北京100081，北京100081，北京100094，保定071000，哈尔滨150030 |
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| 基金项目： | 国家重点基础研究发展计划(973计划);国家高技术研究发展计划(863计划) |
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| 摘 要： | cry8 Ca2基因是本实验室自行分离克隆的一种新型苏云金芽孢杆菌(Bacillus thuringiensis,Bt)杀虫晶体蛋白基因,其表达130 kDa蛋白对铜绿异丽金龟(Anomala corpulenta)和黄褐异丽金龟(A.exoleta)幼虫具有较高的活性。本研究对Cry8Ca2蛋白原毒素及胰凝乳蛋白酶消化片断的纯化条件及活性进行研究,最适的消化条件为质量比1∶20,37℃,消化1 h。通过阴离子交换层析得到纯的活性毒素蛋白。用原毒素与纯化的毒素对铜绿丽金龟幼虫进行生测,Cry8Ca2原毒素蛋白的LC50为0.47μg/g土,纯化的毒素的LC50为0.08μg/g土,毒素的杀虫活性可以达到原毒素的6倍。
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| 关 键 词： | 苏云金芽孢杆菌 Cry8Ca2原毒素 Cry8Ca2毒素 纯化 活性 |
| 修稿时间： | 2007-04-10 |
A study on purification and activity of Cry8Ca2 protein from Bacillus thuringiensis |
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| Wu Hongfu;;Zhang Yancai;Guo Shuyuan;Zhang Jie;Wang Rongyan;Gao Jiguo.A study on purification and activity of Cry8Ca2 protein from Bacillus thuringiensis[J].Plant Protection,2007,33(4):29-32. |
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| Authors: | Wu Hongfu;;Zhang Yancai;Guo Shuyuan;Zhang Jie;Wang Rongyan;Gao Jiguo |
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| Institution: | 1. State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100094, China; 2. Life Science and Technology College, Beijing Institute of Technology, Beijing 100081, China; 3. Instituteof Plant Protection, HebeiAcademy of Agriculture and Forest Sciences, Baoding 071000, China; 4. Life Science College, Northeast Agricultural University, Harbin 150030, China |
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| Abstract: | Cry8Ca2 protein encoded by the gene cry8Ca2 of Bacillus thuringiensis is a kind of insecticidal crystal protein with molecular mass of 130kDa.It is larvicidal to both Anomala corpulenta and A.exoleta.The solubilized Cry8Ca2 protoxin of 130kDa was activated by chymotrypsin at the enzyme / protoxin ratio of 1∶20(w/w).It was further purified by anion exchange chromatography on Resource Q column.The activities of the protoxin and the toxin were evaluated by bioassays.The lethal concentration fifty percent(LC50) of the protoxin was 0.47 μg per gram soil,while that of the toxin was 0.08 μg per gram soil.The activity of the toxins was six folds as that of the protoxin.These results indicate that Cry8Ca2 toxin may be beneficial for biocontrol of both A.corpulenta and A.exoleta. |
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| Keywords: | Bacillus thuringiensis Cry8Ca2 protoxin Cry8Ca2 toxin purification activity |
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