| 苏云金芽胞杆菌cry1Aa19基因的克隆、表达及杀虫活性分析 |
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| 引用本文： | 罗玲,李海涛,刘东明,高继国.苏云金芽胞杆菌cry1Aa19基因的克隆、表达及杀虫活性分析[J].东北农业大学学报,2012,43(1):149-155. |
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| 作者姓名： | 罗玲 李海涛 刘东明 高继国 |
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| 作者单位： | 东北农业大学生命科学学院,哈尔滨,150030 |
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| 基金项目： | 转基因生物新品种培育科技重大专项，植物病虫害生物学国家重点实验室开放基金课题 |
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| 摘 要： | 研究根据cry1Aa类基因的全长序列设计全长引物,以苏云金芽胞杆菌(Bacillus thuringiensis)菌株LS-R-21的基因组DNA为模板扩增出片段大小为3 600 bp的cry1Aa的全长基因,与大肠杆菌(Escherichia coli)表达载体pEB相连接获得含有cry1Aa全长基因的重组质粒pEB-cry1Aa,转入大肠杆菌Rosetta(DE3)菌株中,诱导表达出132 ku的蛋白。该蛋白由1 175个氨基酸组成,其分子质量为132.9964 ku。该基因核苷酸序列已在GenBank中登录,登录号为HQ685121,并且被国际基因命名委员会正式命名为cry1Aa19。杀虫活性测定结果表明,cry1Aa对小菜蛾(Plutella xylostella)有很强的杀虫活性,LC50为1.18μg.mL-1。该基因的获得将为害虫抗性研究及高效工程菌的构建提供了重要的试验材料。
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| 关 键 词： | 苏云金芽胞杆菌 cry1Aa基因 克隆 表达 杀虫活性测定 |
Cloning, expression and insecticidal activity analysis of cry1Aa19 genes from Bacillus thuringiensis |
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| LUO Ling
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LI Haotao
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LIU Dongming
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GAO Jiguo.Cloning, expression and insecticidal activity analysis of cry1Aa19 genes from Bacillus thuringiensis[J].Journal of Northeast Agricultural University,2012,43(1):149-155. |
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| Authors: | LUO Ling LI Haotao LIU Dongming GAO Jiguo |
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| Institution: | (College of Life Sciences,Northeast Agricultural University,Harbin 150030,China) |
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| Abstract: | In this study,the full-length cry1Aa gene of 3600 bp in length was amplified by PCR using the full-length primers were designed according to the complete nucleotide sequences of cry1Aa class gene and genomic DNA of Bacillus thuringiensis strain LS-R-21 as the template,then the cry1Aa gene was cloned into E.coli expression vector pEB,creating the recombinant plasmid pEB-cry1Aa,which was then transformed into Rosetta(DE3),induced a protein with the molecular mass of approximately 132.9964 ku which was consisted by 1 175 amino acids.The sequence was registered in GenBank(Accession Number is HQ685121),and the protein was officially named cry1Aa19 by International Gene Nomenclature Committee.The bioassay results showed that the LC50 of Cry1Aa against Plutella xylostella having strong insecticidal activity was 1.18 μg·mL-1.The gene cry1Aa19 provided an important test material for the study on insecticidal activity and the high effective construc-tion of engineer bacteria. |
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| Keywords: | Bacillus thuringiensis cry1Aa gene clone expression insecticidal activity |
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