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重组真菌细胞色素P450nor2表达条件优化及同源模建分析
发表时间：2010-04-15 浏览量：1675 下载量：499
| 全部作者： | 李书祥，黎晨，袁利玲，李娜，袁永泽，杨江科，闫云君，刘德立 |
| 作者单位： | 华中师范大学生命科学学院, 遗传调控与整合生物学湖北省重点实验室；华中科技大学生命科学与技术学院 |
| 摘 要： | 通过构建重组质粒pET28-p450nor2，成功表达了重组真菌细胞色素P450nor2蛋白。经优化表达条件、提高重组蛋白可溶性大量表达后，采用Ni+-NTA亲和层析分离纯化得到重组蛋白pET-P450nor2. 基于细胞色素P450nor2与同源蛋白P450nor1氨基酸序列的相似性，采用Tripos sybyl软件对细胞色素P450nor2进行同源模建和分子叠合，获得了理想的同源模建结果。经分子动力学优化后，对构建的P450nor2蛋白模型进行模型评价分析。确定模型合理性后，运用FlexX程序对其进行分子对接和功能分析。这将为进一步研究靶酶小分子奠定理论基础。 |
| 关 键 词： | 分子生物学；细胞色素P450nor2；表达条件优化；同源模建分析 |
| Title： | The optimization and purification of the expression of recombinant protein cytochrome P450nor2 and analysis of the homology modeling |
| Author： | LI Shuxiang, LI Chen, YUAN Liling, LI Na, YUAN Yongze, YANG Jiangke, YAN Yunjun, LIU Deli |
| Organization： | College of Life Science, Central China Normal University, Key Laboratory of Pesticide & Chemical Biology of Hubei Province；College of Life Science and Technology, Huazhong University of Science and Technology |
| Abstract： | In order to successfully express the recombinant protein cytochrome P450nor2, recombinant plasmid pET-P450nor2 was constructed. Through optimizing the expression condition of the recombinant protein and using the Ni+-NTA affinity resin column, the soluble and purified protein cytochrome P450nor2 was obtained. By using of the Tripos sybyl software, three-dimensional(3D) model of p450nor2 was established based on high sequence similarity to homologous protein p450nor1 structure and the perfect homology modeling was obtained. The P450nor2 protein model was scored by Procheck process after energy minimization. With the confirmation of the reasonable modeling, molecule docking and function analysis were performed by FlexX. The analysis of function and construction about recombinant protein P450nor2 will lay the solid foundation for the study of small molecules to target enzyme. |
| Key words： | molecular biology; cytochrome P450nor2; optimization of expression conditions; homology modeling |
| 发表期数： | 2010年4月第7期 |
| 引用格式： | 李书祥，黎晨，袁利玲，等. 重组真菌细胞色素P450nor2表达条件优化及同源模建分析[J]. 中国科技论文在线精品论文，2010，3（7）：665-669. |
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