| 吴玉娥1,张杰1,曹景萍1,高继国2,黄大昉3,宋福平1.苏云金芽孢杆菌Cry1Ie1蛋白末端缺失的研究[J].植物保护,2003,29(5):15-19. |
| 苏云金芽孢杆菌Cry1Ie1蛋白末端缺失的研究 |
| Study on truncated Cry1Ie1 protein from Bacillus thuringiensis |
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| DOI： |
| 中文关键词: 基因工程 苏云金芽孢杆菌 杀虫晶体蛋白 Cry1Ie1 小菜蛾 |
| 英文关键词:gene engineering Bacillus thuringiensis insecticidal crystal protein cry1Ie1 Plutella xylostella |
| 基金项目: |
| 吴玉娥1 张杰1 曹景萍1 高继国2 黄大昉3 宋福平1 |
| 1.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;2.东北农业大学 哈尔滨150030;3.中国农业科学院生物技术研究所 北京100081 |
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| 中文摘要: |
| 通过PCR扩增法 ,以cry1Ie1全长基因为模板 ,分别得到不同末端缺失的基因片段 ,转化大肠杆菌BL21(DE3)中诱导表达 ,得到 6种末端缺失蛋白。对小菜蛾生物测定结果表明 ,缺失蛋白IE659、IE656分别缺失了C末端60个和63个氨基酸残基 ,对小菜蛾的杀虫活性与Cry1Ie1全长蛋白相当 ;IE648、IE045分别缺失了C末端 71个氨基酸残基和N末端缺失44个、C末端缺失63个氨基酸残基 ,对小菜蛾的活性提高了50% ;IE633、IE105分别缺失了C末端 86个氨基酸残基和N末端缺失 104个、C末端63个氨基酸残基 ,丧失了对小菜蛾的活性。试验明确Cry1Ie1蛋白的最小活性区N端在45～104氨基酸位点之间 ,C端在633～648氨基酸位点之间。 |
| 英文摘要: |
| With cry1Ie1 gene as template, the different fragments of cry1Ie1 gene were amplfied by PCR method. These fragments were inserted into expressed vector pET-21b, and transformed into Escherichia coli BL21(DE3). Six defferent truncated proteins were induced. The results of bioassays on Plutella xylostella showed that the activities of truncated proteins IE659 (60 amino acid deletion of C-terminus) and IE656 (63 amino acid deletion of C-terminus) against P. xylostella were comparable to that of full-length cry1Ie1 protein. The virulences of IE648 (71 amino acid deletion of C-terminus) and IE045 (44 amino acid deletion of N-terminus and 63 amino acid deletion of C-terminus) against P. xylostella were 50% higher than that of cry1Ie1. However, IE633 (86 amino acid deletion of C-termins) and IE105 (104 amino acid deletion of N-terminus and 63 amino acid deletion of C-terminus) lost the activity to P. xylostella. As the result, it concluded that minimum activity fragment of cry1Ie1 protein was between 45 to 104 amino acid site of N-terminus and 633 to 648 amino acid site of C-terminus. The work laid the foundation of further studies on structure and function interaction of cry1Ie1 gene and provided the theoretic base on constructing engineerring strains and transgenic plants. |
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