大肠杆菌表面展示体系磷酸盐吸附能力的初步研究
关键词：细胞表面展示；正磷酸盐；吸附；污水处理
A preliminary study about the phosphate adsorption capacity of E. coli surface display system
Abstract：In recent years, microbial cell-surface engineering，which uses the microbial cell surface display technology to display foreign proteins on the microbial cell surface，was developed. Cell-surface engineering can be used on cell-catalyst，cell-adsorbent，live vaccine，biosensor and so on，and have a wide application perspective．However, in our country，the microbial cell-surface engineering is still a new research field. In our research, a recombinant Escherichia coli, which displayed phosphate binding protein (PBP) on the cell surface, was used to optimize the cultural and determination conditions. The phosphate binding ability of recombinant strain was further determined by the optimized conditions. The results indicated that recombinant strain showed high efficiency of phosphate binding ability. These results demonstrated that PBP cell surface display system gained significant applications in phosphate removal.
Key words: Cell surface display; orthophospha; adsorption; sewage treatment.
目 录
1前言 6
1.1本课题国内外研究现状概述 6
1.2 有机磷水解酶的大肠杆菌细胞表面展示 6
1.3 重组质粒的构建 7
1.4无机磷酸盐吸附研究进展 7
1.4.1化学除磷法[13-19] 8
1.4.2吸附法除磷 8
1.4.3人工湿地系统 9
1.4.4生物除磷法 9
1.4.5大肠杆菌磷酸盐吸附蛋白 10
1.5本课题的研究目的和意义 11
2 使用的材料试剂、仪器设备和采取的方法、技术 12
2.1 使用的材料试剂、仪器设备 12
2.1.1 材料试剂 12
2.1.2实验试剂 12
2.1.3仪器设备 12
2.2 质粒的提取 13
2.2.1准备工作 13
2.2.2质粒的提取 13
2.3 PCR扩增phoS 13
2.4磷酸盐吸附试验 14
2.4.1 标准曲线的制作 14
2.4.2 磷酸盐浓度的测定 14
2.4.3活菌磷酸盐吸附能力实验 14
2.4.4热致死菌磷酸盐吸附能力试验 15
2.5生长曲线的测定 15
3结果分析 16
3.1 pMB109质粒的提取 16
3.2 PCR扩增phos基因 16
3.3重组质粒的导入对大肠杆菌生长的影响 18
3.4重组菌磷酸盐吸附能力 19
3.4.1磷酸盐标准曲线的测定 19
3.4.2测定磷酸盐吸附能力最适的初始浓度 20
3.4.3绘制重组菌MB109磷酸盐吸附量与时间的关系 21
3.5磷酸盐吸附能力测定 22
4.讨论 24
致谢 25
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