Abdulrahman Wassim, Uhring Muriel, Kolb-Cheynel Isabelle, Garnier Jean-Marie, Moras Dino, Rochel Natacha, Busso Didier, Poterszman Arnaud
Institut de Génétique et de Biologie Moléculaire et Cellulaire, UMR 7104, Illkirch Cedex, France.
Anal Biochem. 2009 Feb 15;385(2):383-5. doi: 10.1016/j.ab.2008.10.044. Epub 2008 Nov 7.
We report a set of baculovirus transfer vectors for parallel expression of proteins in fusion with a panel of affinity tags including GST, protein A, thioredoxin, CBP, and FLAG. This suite includes vectors to generate recombinant baculovirus by homologous recombination in insect cells or using the Bac-to-Bac technology. An application of the vector suite approach to the vitamin D receptor (VDR), a protein mainly expressed as inclusion bodies in Escherichia coli, is presented. We found that expression in fusion with GST and protein A provided an efficient compromise of excellent purification with acceptable yields and costs.
我们报道了一组杆状病毒转移载体，用于与包括谷胱甘肽S-转移酶（GST）、蛋白A、硫氧还蛋白、钙调蛋白（CBP）和FLAG在内的一系列亲和标签融合的蛋白质的平行表达。该套件包括通过昆虫细胞中的同源重组或使用杆状病毒表达系统（Bac-to-Bac技术）产生重组杆状病毒的载体。本文展示了该载体套件方法在维生素D受体（VDR）上的应用，VDR在大肠杆菌中主要以包涵体形式表达。我们发现，与GST和蛋白A融合表达能在出色的纯化效果、可接受的产量和成本之间实现有效平衡。
