大学化学 >> 2022, Vol. 37 >> Issue (12): 2112046.doi: 10.3866/PKU.DXHX202112046

化学实验 上一篇    下一篇

金属β-内酰胺酶L1的制备和动力学表征线上线下混合实验

和媛(), 张烨, 王梦琛, 崔斌, 李延, 樊海明()   

  • 收稿日期:2021-12-17 录用日期:2022-03-10 发布日期:2022-03-25
  • 通讯作者: 和媛,樊海明 E-mail:yuanhe@nwu.edu.cn;fanhm@nwu.edu.cn
  • 作者简介:樊海明, Email: fanhm@nwu.edu.cn
    和媛, Email: yuanhe@nwu.edu.cn
  • 基金资助:
    中国高等教育学会理科教育专业委员会研究课题(21ZSLKJYYB13);陕西省高等教育学会高等教育科学研究项目(XGH21083);教指委教学研究与实践项目(H20210602);教指委教学研究与实践项目(51408205);陕西省一流专业建设项目;西北大学本科人才培养建设项目(XM05190493);西北大学本科人才培养建设项目(XM05190808)

Preparation and Kinetic Characterization of Metallo-β-Lactamase L1 in an Online-Offline Blended Teaching Mode

Yuan He(), Ye Zhang, Mengchen Wang, Bin Cui, Yan Li, Haiming Fan()   

  • Received:2021-12-17 Accepted:2022-03-10 Published:2022-03-25
  • Contact: Yuan He,Haiming Fan E-mail:yuanhe@nwu.edu.cn;fanhm@nwu.edu.cn

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摘要：

蛋白质的基因克隆与表达是许多化学生物学实验的重要内容，但因原理概念抽象、操作步骤多、耗时长的缺点限制了其在常规本科实验教学中的有效开展。基于本科化学生物学创新实验“金属β-内酰胺酶L1的制备和动力学表征”，对原实验教学内容与教学方式进行了创新性改进。主要改进包括：(1) 利用移动设备进行金属β-内酰胺酶L1的制备及表征线上虚拟仿真实验，含聚合酶链反应扩增目的基因、重组质粒转化感受态细胞、蛋白诱导表达等10个模块。线上高仿真实验场景和人机交互性操作给学生提供高度沉浸感实验体验，集中连贯的实验教学使知识构架更清晰、更完整。(2) 针对重点实验及拓展实验进行线下课堂操作，新增等温滴定微量热法表征酶活线下拓展实验。该混合教学模式实现了虚实结合、线上线下融合，既拓宽了学生的视野，培养了动手能力和创新意识，也提高了教学质量和课程管理，在实践中取得了较好的反馈，可为其他院校本科实验教学提供参考。

关键词: 金属β-内酰胺酶L1, 酶促反应动力学, 等温滴定微量热法, 虚拟仿真实验, 线上线下混合教学

Abstract:

Recombinant protein preparation is an important part of many chemical biology experiments, but its effective implementation in a regular undergraduate experiment course is limited by the long period of time required. This work reports innovative improvements on the content and teaching methods in an undergraduate teaching experiment: "Preparation and kinetic characterization of metallo-β-lactamase L1". The main improvements include: (1) using online virtual simulation experiments on a mobile device to study the whole process of L1 preparation and conduct a kinetics study; and (2) offline classroom operation for compulsory and extended experiments. The online resource contains 10 modules, such as polymerase chain reaction of a target gene, recombinant plasmid transformation, protein induction, etc. High-quality simulation of experimental scenarios and human-machine interactive operations provide students with a highly immersive experience, and the coherent experimental arrangement presents a clear knowledge framework. A new offline extension experiment was added to enable the use of the isothermal titration microcalorimetric method to characterize the kinetic parameters of the L1 enzyme. This blended teaching mode realizes the integration of online and offline teaching, which can not only improve the quality and efficiency of teaching, but also broaden students' horizons and cultivate their hands-on ability and innovation awareness. Thus far, good feedback has been received from preliminary implementation, which is expected to provide a reference for undergraduate experimental teaching in other institutions.

Key words: Metallo-β-lactamase L1, Kinetics of enzymatic reaction, Isothermal titration microcalorimetry, Virtual simulation experiment, Online-offline blended teaching