| 金瑾,杨少媛,朱嘉,雷振,郑积敏,贾宗超.化学通报,2015,78(1):85-88. |
| 大肠杆菌MinCD蛋白复合物纯化方法和结晶筛选 |
| Purification and Crystallization of MinCD Complex from Escherichia coli |
| 投稿时间：2014-05-23 修订日期：2014-06-20 |
| DOI： |
| 中文关键词: 细胞分裂 复合物 结晶 MinC MinD |
| 英文关键词:Cell division Complex Crystallization MinC MinD |
| 基金项目: |
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| 中文摘要: |
| 大肠杆菌在细胞分裂时, 细胞中部潜在分裂位点的选择受到Min系统（MinC、MinD和MinE蛋白）的精确调控, 其中MinC与MinD可以形成复合物抑制Z分裂环在错误的位置形成。本研究采用基因克隆方法获得minC、minD基因的克隆，并表达得到MinC和MinD的重组蛋白，并用亲和层析与凝胶过滤层析对所得蛋白进行纯化，通过minC、minD分别单独转化、表达纯化后混合和minC、minD共转化法两种方法得到MinCD蛋白复合物，并分别对其进行晶体筛选。结果表明，共转化法得到的MinCD复合物纯度较好，且比例约为2：1，并通过晶体筛选初步获得针状蛋白晶体，为MinCD复合物的结构解析提供了实验基础。 |
| 英文摘要: |
| In Escherichia coli, the mid-cell division site is spatially regulated by Min system which including protein MinC、MinD and MinE. MinC and MinD can form complex to inhibit the formation of the septum Z ring at wrong site. In this study, minC、minD genes were constructed into vector to express recombination proteins, then MinC and MinD proteins were purified by affinity chromatography and gel filtration chromatography. Two different methods, binding MinC with MinD after being expressed and purified respectively, and co-transformation, co-expression and co-purification of MinC and MinD were tried to gain the crystal of MinCD complex for further structural analysis. The results show that through co-transformation method, purified MinCD complex were obtained in proportion about 2:1 and needle shape crystals of MinCD complex was observed after crystal screening. This study provide basic method for structure determination of MinCD complex. |
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