An Adaptable Investigative Graduate Laboratory Course for Teaching Protein Purification
An Adaptable Investigative Graduate Laboratory Course for Teaching Protein Purification
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DOI：
10.1002/bmb.20827
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发表时间：
2014-11-01
影响因子：
1.4
通讯作者：
Keller, Lani C.
中科院分区：
文献类型：
--
作者：
Carroll, Christopher W.;Keller, Lani C.
这门适应性强的研究生实验课程为学生提供了探索广泛技术的机会，同时允许讲师自由地结合他们自己的个人研究兴趣。课程设计包括在一个学期内完成两个顺序的净化方案。第一部分包括大肠杆菌中重组gfp结合蛋白的表达和纯化。学生纯化的gfp结合蛋白随后在课程的第二部分使用，从细胞或组织提取物中免疫沉淀gfp标记的蛋白及其潜在的相互作用伙伴。作为一个例子，我们描述了从黑腹果蝇幼虫提取物中提取的gfp标记蛋白的免疫沉淀，这些蛋白与人类疾病相关的蛋白同源，然后用western blotting来检查学生的实验结果。然而，gfp融合蛋白在不同生物中的广泛应用使研究人员能够根据他们的具体研究计划定制课程的第二部分，同时保持灵活性，使学生参与主动学习。学生的评价显示出他们对研究的真正热情，对所使用的技术及其背后的理论都有深入的了解。(c) 2014.国际生物化学与分子生物学联合会，42(6):486-494,2014。
This adaptable graduate laboratory course on protein purification offers students the opportunity to explore a wide range of techniques while allowing the instructor the freedom to incorporate their own personal research interests. The course design involves two sequential purification schemes performed in a single semester. The first part comprises the expression and purification of a recombinant GFP-binding protein from E. coli. The student-purified GFP-binding protein is then used in the second part of the course to immunoprecipitate GFP-tagged proteins, and their potential interacting partners, from cell or tissue extracts. As an example, we describe the immunoprecipitation of GFP-tagged proteins from Drosophila melanogaster larval extracts that are homologous to proteins implicated in human diseases, followed by western blotting to examine student experimental outcomes. However, the widespread availability of GFP-fusion proteins in diverse organisms enables researchers to tailor the second part of the course to their specific research programs while maintaining the flexibility to engage students in active learning. Student evaluations indicate a genuine excitement for research and in depth knowledge of both the techniques performed and the theory behind them. (c) 2014 by The International Union of Biochemistry and Molecular Biology, 42(6):486-494, 2014.