基于PAMAM-Au复合纳米颗粒的四环素及其差向异构体的受体分析方法研究

项目名称： 基于PAMAM-Au复合纳米颗粒的四环素及其差向异构体的受体分析方法研究

项目编号： No.31502118

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 农业科学

项目作者： 曾昆

作者单位： 江苏大学

项目金额： 21万元

中文摘要： 四环素类药物（Tetracyclines，TCs）的过量使用导致其在动物源性食品中的广泛残留。TCs经动物体内代谢后可转化为毒性更强的差向异构体，TCs及其差向异构体的残留对动物源性食品安全造成了极大的威胁。为监控动物源性食品TCs残留，四种TCs（四环素、土霉素、金霉素和多西环素）及其三种差向异构体（除多西环素）被列为残留标志物，因此建立针对此四种TCs及三种差向异构体的高效、快速的分析方法具有重要意义。本项目以上四种TCs及三种差向异构体为研究对象，体外克隆并表达四环素阻遏蛋白（Tet repressor, TetR）；合成聚酰胺-胺（Polyamidoamine, PAMAM），修饰纳米金颗粒；TetR共价偶联PAMAM-Au；进而构建广谱、灵敏的分析方法，实现四环素类及其差向异构体的快速、准确测定。本项目的成功实施为药物多残留分析提供新的思路，并为我国食品安全提供可靠的的技术支持。

中文关键词： 四环素；异构体；四环素阻遏蛋白；PAMAM；复合纳米颗粒

英文摘要： Tetracycline antibiotics (TCs) are widely used as veterinary drugs for food-producing animals because of their broad-spectrum activity and cost effectiveness. TCs residues can cause toxic or hypersensitive reactions in sensitive individuals and also transfer of drug-resistant bacteria from food to humans. After metabolized in the body, TCs can be transformed to their 4-epimers, which have lower activity and higher toxic effect. According to European Community, the residue of interest is the sum of the parent compounds TCs (Tetracycline, Tetracycline antibiotics (TCs) are widely used as veterinary drugs for food-producing animals because of their broad-spectrum activity and cost effectiveness. TCs residues can cause toxic or hypersensitive reactions in sensitive individuals and also transfer of drug-resistant bacteria from food to humans. After metabolized in the body, TCs can be transformed to their 4-epimers, which have lower activity and higher toxic effect. According to European Community, the residue of interest is the sum of the parent compounds TCs (Tetracycline, Oxytetracycline, Chlortetracycline and Doxycycline) and their 4-epimers, except for doxycycline, where only the parent compound is included in the MRL. None of rapid assay reported could detect all these TCs and their 4-epimers. In this study, we try to clone and express the binding region of Tet repressor through prokaryotic expression in E. Coli. NH2-terminated Polyamidoamine (PAMAM) are synthesized and characterization and PAMAM-Au nanocomposites are developed using reduction method. TetR are linked to NH2 end in the surface of PAMAM-Au. With PAMAM-Au-TetR, a new broad-spectrum assay is developed to detect TCs and their 4-epimers in milk. The implement of the project should be to explore new recognized molecules and detection techniques of multi-residue to provide technical support for food security.

英文关键词： Tetracycline; Epimer;Tet repressor;PAMAM;nanocomposites