Investigating the Determinants of Substrate Binding through a Semester-Long, Project-Oriented Biochemistry Laboratory Course,Journal of Chemical Education

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Investigating the Determinants of Substrate Binding through a Semester-Long, Project-Oriented Biochemistry Laboratory Course
Journal of Chemical Education ( IF 2.5 ) Pub Date : 2018-08-23 00:00:00 , DOI: 10.1021/acs.jchemed.8b00028
Catherine A. Sarisky ₁ , Timothy W. Johann ₁

Affiliation

A semester-long hypothesis-driven laboratory project for second-semester biochemistry has been developed. Working independently, students propose a hypothesis about the role of one amino acid residue in the active site of 5,10-methenyltetrahydrofolate synthetase (MTHFS). They then test this hypothesis by site-directed mutagenesis of a plasmid encoding the wild-type enzyme, overexpression and purification of the mutant protein, and detailed kinetic characterization. Student success in this laboratory course requires attention to detail and careful preservation of materials from week to week. Controls are carefully selected to allow diagnosis of experimental failures, and students are required to discuss them in their laboratory reports. The combination of independent work coupled with experiments in which results from one week influence the next provides students with a model of professional research activities that is absent from traditional laboratory experiments.

中文翻译：

通过为期半学期，面向项目的生化实验课程研究底物结合的决定因素

已经开发了一个学期长的假设驱动的第二学期生物化学实验室项目。学生们独立工作，提出了一个氨基酸残基在5,10-亚甲基四氢叶酸合成酶（MTHFS）活性位点中的作用的假设。然后，他们通过对编码野生型酶的质粒进行定点诱变，突变蛋白的过表达和纯化以及详细的动力学表征，来检验这一假设。学生要在本实验室课程中取得成功，需要每周注意细节并仔细保存材料。仔细选择控件以诊断实验失败，要求学生在实验室报告中进行讨论。

更新日期：2018-08-23

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