| 赤翅甲抗冻蛋白基因的原核表达及蛋白生物活性检测 |
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| 引用本文： | LIU Zhong-Yuan,张富春,王芸,吕国栋.赤翅甲抗冻蛋白基因的原核表达及蛋白生物活性检测[J].昆虫学报,2005,48(2):179-183. |
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| 作者姓名： | LIU Zhong-Yuan 张富春 王芸 吕国栋 |
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| 作者单位： | 新疆大学生命科学与技术学院分子生物学重点实验室，新疆生物资源基因工程重点实验室 |
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| 基金项目： | 国家科技攻关项目 (2001BA901A32) |
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| 摘 要： | 根据GenBank中序列人工合成赤翅甲Dendroides canadensis的抗冻蛋白基因（afp），将其克隆到载体pGEX-4T-1上，构建融合表达的重组质粒，转化大肠杆菌 BL21并进行原核表达。通过优化表达的诱导条件和SDS-PAGE检测，证明人工合成的赤翅甲抗冻蛋白基因能够特异性地表达，并以可溶性融合蛋白形式存在，相对分子质量约为40 kD。抗冻蛋白的生物活性检测表明，赤翅甲的抗冻融合蛋白能够提高细菌的耐寒能力。
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| 关 键 词： | 赤翅甲 抗冻蛋白 原核表达 抗冻活性 |
| 文章编号： | 0454-6296(2005)02-0179-05 |
| 修稿时间： | 2004年1月4日 |
Expression of the antifreeze protein gene of a pyrochroid beetle Dendroides canadensis in prokaryote and detection of the protein biological activity |
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| LIU Zhong-Yuan,ZHANG Fu-Chun,WANG Yun,LU Guo-Dong.Expression of the antifreeze protein gene of a pyrochroid beetle Dendroides canadensis in prokaryote and detection of the protein biological activity[J].Acta Entomologica Sinica,2005,48(2):179-183. |
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| Authors: | LIU Zhong-Yuan ZHANG Fu-Chun WANG Yun LU Guo-Dong |
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| Affiliation: | Key Laboratory of Molecular Biology, College of Life Science and Technology |
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| Abstract: | In order to express a pyrochroid beetle Dendroides canadensis antifreeze protein in E. coli BL21 and to investigate the properties of recombinant antifreeze protein, the D. canadensis afp gene was synthesized, cloned into the pGEX-4T-1, and transformed into E. coli BL21. The optimal inductive condition was selected by different experimental approaches. The results of SDS-PAGE analysis indicated that the antifreeze protein was expressed successfully in E.coli BL21. The expressed antifreeze protein, existing in the soluble and fusion form, with the molecular weight of about 40 kD, could increase low temperature resistance of bacteria as shown by the biological activity detection. |
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| Keywords: | Dendroides canadensis antifreeze protein prokaryotic expression biological activity |
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