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DOI: 10.3791/55300-v
Keith C. Heyde*1,2, Felicia Y. Scott*3, Sung-Ho Paek3, Ruihua Zhang3, Warren C. Ruder3,4
1Department of Mechanical Engineering,Carnegie Mellon University, 2Engineering Science and Mechanics Program,Virginia Polytechnic Institute and State University, 3Department of Biological Systems Engineering,Virginia Polytechnic Institute and State University, 4Department of Bioengineering,University of Pittsburgh
Please note that some of the translations on this page are AI generated. Click here for the English version.
本文提出了一系列的开发改造的细胞和,使综合工程大肠杆菌控制和操纵可编程材料的表面功能化表面的协议。
这些程序的总体目标是通过结合基因改造和表面功能化策略,使用合成工程大肠杆菌来控制和纵可编程材料表面。这种方法可以帮助回答分子医学和环境监测等领域的关键问题。通过使用基因编程细胞来解释局部环境并相应地修改功能化材料,我们为各种应用提供了模块化工具。
这种技术的主要优点是活细胞能够充当动态传感器,能够通过功能化接口读取、处理和记录周围的条件。根据文本方案从产生生物素的大肠杆菌中制备溶液并收集富含生物素的上清液后,将 1.4 微升 SPDP 溶液添加到 20 微升链霉亲和素或 SA 溶液中。将试管包裹在铝箔中,并在室温下孵育一个半小时,以使 SPDP 交联剂通过氨基与 SA 键合,形成吡啶基二硫代活化的 SA。孵育后,向试管中加入 2.4 μL DTT 溶液,并在室温下孵育样品 1 小时,以允许吡啶-2-硫酮裂解,从而产生巯基活化的 SA。接下来,将 7.5 微升 SCC 溶液加入 72 微升 HRP 溶液中,用铝箔包裹,并在室温下孵育一个半小时。
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