| 苏云金芽孢杆菌Rpp02菌株cry1Ac20基因的克隆及表达 |
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| 引用本文： | 谭芙蓉,李平,郑爱萍,周华强,郑秀丽.苏云金芽孢杆菌Rpp02菌株cry1Ac20基因的克隆及表达[J].昆虫学报,2006,49(6):950-954. |
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| 作者姓名： | 谭芙蓉 李平 郑爱萍 周华强 郑秀丽 |
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| 作者单位： | (四川农业大学水稻研究所，四川温江611130) |
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| 基金项目： | 国家“863”高技术研究发展计划项目(2002AA212151)，教育部长江学者和创新团队发展计划(IRT0453) |
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| 摘 要： | Rpp02菌株是本实验室分离的一株对鳞翅目等多种害虫具有高毒力的苏云金芽孢杆菌莫里逊亚种 (Bacillus thuringiensis subsp. morrisoni), 经PCR检测，它含有cry1Ac基因。对其基因组DNA进行PCR扩增，得到大约4 kb的产物。测序结果表明，该片段含有一个较大的ORF框，基因编码区为3 534 bp，编码1 177个氨基酸，分子量为133.144 kD，pI 4.952, 为弱酸性蛋白质，亮氨酸（Leu）、丝氨酸（Ser）、谷氨酸（Glu）3种氨基酸含量最高，分别为8.0%、7.8%、7.7%。该基因序列与cry1Ac序列同源性达到99%，并被国际Bt杀虫晶体蛋白基因命名委员会命名为cry1Ac20。生物测定表明，该基因在大肠杆菌中得到了表达，表达产物具有较强的杀虫效果，试喂菜青虫48 h后,校正死亡率为88.78%。
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| 关 键 词： | 苏云金芽孢杆菌 cry1Ac20基因 克隆 表达 菜青虫 |
| 文章编号： | 0454-6296(2006)06-0950-05 |
| 收稿时间： | 12 28 2005 12:00AM |
| 修稿时间： | 05 30 2006 12:00AM |
Cloning and expression of cry1Ac20 gene from Bacillus thuringiensis strain Rpp02 |
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| TAN Fu-Rong,LI Ping,ZHENG Ai-Ping,ZHOU Hua-Qiang,ZHENG Xiu-Li.Cloning and expression of cry1Ac20 gene from Bacillus thuringiensis strain Rpp02[J].Acta Entomologica Sinica,2006,49(6):950-954. |
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| Authors: | TAN Fu-Rong LI Ping ZHENG Ai-Ping ZHOU Hua-Qiang ZHENG Xiu-Li |
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| Affiliation: | (Rice Research Institute, Sichuan Agricultural University, Wenjiang, Sichuan 611130, China) |
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| Abstract: | Bacillus thuringiensis subsp.morrisoni strain Rpp02 isolated by researchers in our laboratory was highly toxic to several kinds of insect pests.Results of PCR analysis indicated that strain Rpp02 contained the cry1Ac gene,and subsequently a novel cry1Ac gene was cloned from this strain by PCR techniques.The results of sequence analysis showed that the novel gene cry1Ac20,named by B.thuringiensis Pesticidal Crystal Protein Nomenclature Committee,contained an open reading frame of 3 534 nucleotides encoding a protein of 1 177 amino acids with a predicted molecular mass of 133.144 kDa and isoelectric point of 4.952.Compared with other known cry1Ac genes,cry1Ac20 has shown as high as 99% nucleotide sequence homology.Bioassay showed that the toxic protein appeared high insecticidal activity against Pieris rapae L.,and the corrected mortality was 88.78% 48 h after feeding with leaves immersed in cultures containing transformants. |
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| Keywords: | Bacillus thuringiensis cry1Ac20 gene cloning expression |
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