A set of baculovirus transfer vectors for screening of affinity tags and parallel expression strategies

被引:56
作者
Abdulrahman, Wassim [1 ]
Uhring, Muriel [1 ]
Kolb-Cheynel, Isabelle [1 ]
Garnier, Jean-Marie [1 ]
Moras, Dino [1 ]
Rochel, Natacha [1 ]
Busso, Didier [1 ]
Potersman, Arnaud [1 ]
机构
[1] Inst Genet & Biol Mol & Cellulaire, UMR 7104, F-67404 Illkirch Graffenstaden, France
关键词
PROTEIN EXPRESSION; ESCHERICHIA-COLI; SYSTEM; THROUGHPUT; INSECT; CELLS;
D O I
10.1016/j.ab.2008.10.044
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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. (C) 2008 Elsevier Inc. All rights reserved.
引用
收藏
页码:383 / 385
页数:3
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共 14 条
  • [11] Time reduction and process optimization of the baculovirus expression system for more efficient recombinant protein production in insect cells
    Philipps, B
    Rotmann, D
    Wicki, M
    Mayr, LM
    Forstner, M
    [J]. PROTEIN EXPRESSION AND PURIFICATION, 2005, 42 (01) : 211 - 218
  • [12] Functional and structural characterization of the insertion region in the ligand binding domain of the vitamin D nuclear receptor
    Rochel, N
    Tocchini-Valentini, G
    Egea, PF
    Juntunen, K
    Garnier, JM
    Vihko, P
    Moras, D
    [J]. EUROPEAN JOURNAL OF BIOCHEMISTRY, 2001, 268 (04): : 971 - 979
  • [13] Making the most of affinity tags
    Waugh, DS
    [J]. TRENDS IN BIOTECHNOLOGY, 2005, 23 (06) : 316 - 320
  • [14] Improving baculovirus recombination
    Zhao, YG
    Chapman, DAG
    Jones, IM
    [J]. NUCLEIC ACIDS RESEARCH, 2003, 31 (02) : E6 - 6