Wheat germ systems for cell-free protein expression

被引:84
|
作者
Harbers, Matthias [1 ,2 ]
机构
[1] RIKEN, Ctr Life Sci Technol, Tsurumi ku, Yokohama, Kanagawa 2300045, Japan
[2] CellFree Sci, Tsurumi ku, Yokohama, Kanagawa 2300046, Japan
关键词
Wheat germ; Cell-free; Protein expression; Automation; Isotopic labeling; Unnatural amino acid; HIGH-THROUGHPUT CLONING; FREE TRANSLATION; AMINO-ACIDS; FUNCTIONAL-ANALYSIS; MASS-SPECTROMETRY; HIGHLY EFFICIENT; MESSENGER-RNA; PURIFICATION; LIBRARY; ROBUST;
D O I
10.1016/j.febslet.2014.05.061
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Cell-free protein expression plays an important role in biochemical research. However, only recent developments led to new methods to rapidly synthesize preparative amounts of protein that make cell-free protein expression an attractive alternative to cell-based methods. In particular the wheat germ system provides the highest translation efficiency among eukaryotic cell-free protein expression approaches and has a very high success rate for the expression of soluble proteins of good quality. As an open in vitro method, the wheat germ system is a preferable choice for many applications in protein research including options for protein labeling and the expression of difficult-to-express proteins like membrane proteins and multiple protein complexes. Here I describe wheat germ cell-free protein expression systems and give examples how they have been used in genome-wide expression studies, preparation of labeled proteins for structural genomics and protein mass spectroscopy, automated protein synthesis, and screening of enzymatic activities. Future directions for the use of cell-free expression methods are discussed. (C) 2014 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
引用
收藏
页码:2762 / 2773
页数:12
相关论文
共 50 条
  • [1] A novel family of expression vectors with multiple affinity tags for wheat germ cell-free protein expression
    Nagy, Szilvia Krisztina
    Kallai, Brigitta Margit
    Andras, Judit
    Meszaros, Tamas
    BMC BIOTECHNOLOGY, 2020, 20 (01)
  • [2] A novel family of expression vectors with multiple affinity tags for wheat germ cell-free protein expression
    Szilvia Krisztina Nagy
    Brigitta Margit Kállai
    Judit András
    Tamás Mészáros
    BMC Biotechnology, 20
  • [3] The Wheat-Germ Cell-Free Expression System
    Takai, Kazuyuki
    Sawasaki, Tatsuya
    Endo, Yaeta
    CURRENT PHARMACEUTICAL BIOTECHNOLOGY, 2010, 11 (03) : 272 - 278
  • [4] Wheat germ cell-free platform for eukaryotic protein production
    Vinarov, Dmitriy A.
    Loushin Newman, Carrie L.
    Markley, John L.
    FEBS JOURNAL, 2006, 273 (18) : 4160 - 4169
  • [5] The growing impact of lyophilized cell-free protein expression systems
    Hunt, J. Porter
    Yang, Seung Ook
    Wilding, Kristen M.
    Bundy, Bradley C.
    BIOENGINEERED, 2017, 8 (04) : 325 - 330
  • [6] A wheat germ cell-free system is a novel way to screen protein folding and function
    Morita, EH
    Sawasaki, T
    Tanaka, R
    Endo, Y
    Kohno, T
    PROTEIN SCIENCE, 2003, 12 (06) : 1216 - 1221
  • [7] Phosphorylation and Alternative Translation on Wheat Germ Cell-Free Protein Synthesis of the DHBV Large Envelope Protein
    David, Guillaume
    Fogeron, Marie-Laure
    Montserret, Roland
    Lecoq, Lauriane
    Page, Adeline
    Delolme, Frederic
    Nassal, Michael
    Bockmann, Anja
    FRONTIERS IN MOLECULAR BIOSCIENCES, 2019, 6
  • [8] Wheat germ cell-free expression system as a pathway to improve protein yield and solubility for the SSGCID pipeline
    Guild, Katherine
    Zhang, Yang
    Stacy, Robin
    Mundt, Elizabeth
    Benbow, Sarah
    Green, Amanda
    Myler, Peter J.
    ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS, 2011, 67 : 1027 - 1031
  • [9] Easy Synthesis of Complex Biomolecular Assemblies: Wheat Germ Cell-Free Protein Expression in Structural Biology
    Fogeron, Marie-Laure
    Lecoq, Lauriane
    Cole, Laura
    Harbers, Matthias
    Bockmann, Anja
    FRONTIERS IN MOLECULAR BIOSCIENCES, 2021, 8
  • [10] Formation of circular polyribosomes in wheat germ cell-free protein synthesis system
    Madin, K
    Sawasaki, T
    Kamura, N
    Takai, K
    Ogasawara, T
    Yazaki, K
    Takei, T
    Miura, KI
    Endo, Y
    FEBS LETTERS, 2004, 562 (1-3) : 155 - 159