Johnson Josiah W, Mitchell Christian D, Deloach Anna M, Simpson Hannah E, Dunlap Tori B
Department of Chemistry, University of Central Arkansas, 201 Donaghey Avenue, Conway, Arkansas 72035, United States.
J Chem Educ. 2019 Oct 1;96(11):2606-2610. doi: 10.1021/acs.jchemed.8b00674. Epub 2019 Nov 12.
Protein structure, function, and signaling are a large portion of biochemistry. Because of this, proteins are often used as model systems in biochemistry laboratory courses, where a course-long project might comprise protein expression, purification, and characterization. Two common protein expression methods are isopropyl -d-1-thiogalactopyranoside (IPTG) induction, which utilizes easy-to-make media but requires extensive cell-growth monitoring that is time-intensive, and autoinduction, which employs multicomponent media that are time-consuming to make but require no cell-growth monitoring. A protein expression method that is a hybrid of IPTG induction and autoinduction is presented. The hybrid method utilizes the medium of IPTG induction and the no-cell-growth-monitoring induction process of autoinduction, saving hands-on time in the protein expression phase to allow more time for protein characterization while still having students execute each step.
蛋白质结构、功能及信号传导是生物化学的重要组成部分。因此，蛋白质常被用作生物化学实验课程中的模型系统，在这类课程中，一个贯穿整门课程的项目可能包括蛋白质表达、纯化及特性分析。两种常见的蛋白质表达方法分别是异丙基-β-D-硫代半乳糖苷（IPTG）诱导法，该方法使用易于制备的培养基，但需要对细胞生长进行大量耗时的监测；以及自诱导法，该方法采用多组分培养基，制备过程耗时，但无需监测细胞生长。本文介绍了一种结合了IPTG诱导法和自诱导法的蛋白质表达方法。这种混合方法采用IPTG诱导法的培养基以及自诱导法中无需监测细胞生长的诱导过程，节省了蛋白质表达阶段的实际操作时间，以便有更多时间进行蛋白质特性分析，同时仍让学生执行每个步骤。
