模拟酶的纯化,作为生物化学入门课程中的“干”实践

José Carlos Cameselle, Alicia Cabezas, José Canales, Marı́a Jesús Costas, Ángeles Faraldo, Ascensión Fernández, Rosa Marı́a Pinto, João Meireles Ribeiro
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引用次数: 0

摘要

这篇文章描述了一个“干实验室”的实践,其中多步骤纯化酶是模拟。它被设计用于初级生物化学入门课程的本科生,目的是让他们对复杂的研究型实验所涉及的努力强度有一个粗略的了解。模拟提纯步骤为:制备肝溶萃取物、硫酸铵分馏、凝胶过滤、离子交换和染料配体亲和层析。在模拟之前,我们的学生通过简短的“食谱”实验室实验,在某些情况下,特别演示,熟悉所涉及的准备和分析技术。对于模拟,学生将获得详细的准备和分析实验方案,以及获得的原始数值或图形数据。他们必须进行必要的计算和绘图以生成净化表。实施实践所需的27页学生手册由作者提供给感兴趣的教师,作为可打印的电子文件。
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The simulated purification of an enzyme as a `dry’ practical within an introductory course of biochemistry

This article describes a `dry-laboratory’ practical in which the multi-step purification of an enzyme is simulated. It has been devised to be implemented with beginner undergraduates taking an introductory course in biochemistry, with the aim of giving them a glimpse of the intensity of effort involved in complex, research-oriented experiments. The purification steps simulated are: preparation of a liver soluble extract, ammonium sulfate fractionation, gel filtration, ion-exchange and dye-ligand affinity chromatography. Before the simulation, our students are familiar with the preparative and analytical techniques involved, through short `cookbook’ laboratory experiments and, in some cases, ad hoc demonstrations. For the simulation, the students are given detailed protocols of the preparative and analytical experiments, and the raw numeric or graphical data obtained. They have to perform the calculations and graphing necessary to produce a purification table. The 27-page Student Booklet needed to implement the practical is offered by the authors to interested teachers, as a printable electronic file.

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