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用于核磁共振的昆虫细胞中同位素标记蛋白质的生产。

Production of isotope-labeled proteins in insect cells for NMR.

作者信息

Franke Bastian, Opitz Christian, Isogai Shin, Grahl Anne, Delgado Leonildo, Gossert Alvar D, Grzesiek Stephan

机构信息

Focal Area Structural Biology and Biophysics, Biozentrum, University of Basel, 4056, Basel, Switzerland.

Department of Biology, ETH Zürich, 8093, Zürich, Switzerland.

出版信息

J Biomol NMR. 2018 Jul;71(3):173-184. doi: 10.1007/s10858-018-0172-7. Epub 2018 Apr 23.

Abstract

Baculovirus-infected insect cells have become a powerful tool to express recombinant proteins for structural and functional studies by NMR spectroscopy. This article provides an introduction into the insect cell/baculovirus expression system and its use for the production of recombinant isotope-labeled proteins. We discuss recent advances in inexpensive isotope-labeling methods using labeled algal or yeast extracts as the amino acid source and give examples of advanced NMR applications for proteins, which have become accessible by this eukaryotic expression host.

摘要

杆状病毒感染的昆虫细胞已成为通过核磁共振光谱进行结构和功能研究来表达重组蛋白的强大工具。本文介绍了昆虫细胞/杆状病毒表达系统及其在生产重组同位素标记蛋白中的应用。我们讨论了使用标记的藻类或酵母提取物作为氨基酸来源的廉价同位素标记方法的最新进展,并给出了通过这种真核表达宿主得以实现的蛋白质高级核磁共振应用实例。

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