| 苏云金芽孢杆菌营养期杀虫蛋白基因vip3A的研究 |
| |
| 引用本文： | 刘荣梅,张杰,高继国,宋福平.苏云金芽孢杆菌营养期杀虫蛋白基因vip3A的研究[J].高技术通讯,2004,14(9):39-42. |
| |
| 作者姓名： | 刘荣梅 张杰 高继国 宋福平 |
| |
| 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,北京,100094;东北农业大学生命科学院,哈尔滨,150030;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,北京,100094;东北农业大学生命科学院,哈尔滨,150030 |
| |
| 基金项目： | 863计划 ( 2 0 0 1AA2 14 0 11)，973规划 ( 2 0 0 3CB114 2 0 1)资助项目 |
| |
| 摘 要： | 根据已知vip3A基因序列设计一对特异性引物VIP3／VIP5，对218株Bt菌株进行PCR鉴定，有51株含有vip3A类基因，其中12株为不同亚种的标准菌株，有4株标准菌株不含有该基因。从Bt C9菌株中分离克隆了vip—C9基因，并插入表达载体pET—21b，转化大肠杆菌BL21，诱导表达出分子量88．6kDa的可溶性蛋白，表达产物对甜菜夜蛾和棉铃虫具有较高的杀虫活性，LC50分别为0．42ng／mg和25．95ng／mg，对小菜蛾活性较低。构建了缺失蛋白Vip—C9—N(N端去除39个氨基酸组成的信号肽序列)，杀虫活性测定结果表明其对甜菜夜蛾的活性显著降低，说明N端39个氨基酸对甜菜夜蛾的活性是必需的。
|
| 关 键 词： | 苏云金芽孢杆菌 营养期杀虫蛋白 甜菜夜蛾 标准菌株 |
The Research on vip3A Genes from Bacillus thuriniensis Strains |
| |
| Abstract: | A pair of specific primers was designed according to the sequences of known vip3A genes. The vip3A-type genes were assayed by PCR amplification for 218 strains. 39 strains and 12 standard strains contained vip3A genes. No vip3A gene was found in 4 standard strains. A vip3A gene named vip-C9 was cloned from Bt strain Bt C9 and expressed normally in BL21strain using expression vector pET-21b. The soluble product with molecular weight of 88.6 kDa was induced and highly toxic to Spodoptera exigua and Helicoverpa armigera larvae with LC 50 of 0.42ng/mg and 25.59ng/mg respectively, while it showed low toxic to Plutella xylostella larvae. The truncation protein Vip-C9-N was constructed by deletion of N-terminal signal peptide sequence consisting of 39 amino acid residues. The activity of Vip-C9-N to Spodoptera exigua larvae significantly decreased. The results showed that N-terminal signal peptide was necessary for activity of Vip-C9 against Spodoptera exigua. |
| |
| Keywords: | Bacillus thuriniensis Vegetative insecticidal proteins Spodoptera exigua Standard strains |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ |
