Multiple inhibitory factor removal from an Escherichia coli cell extract improves cell-free protein synthesis(ENZYMOLOGY, PROTEIN ENGINEERING, AND ENZYME TECHNOLOGY)

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概要

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• Cell-free protein synthesis systems are generally influenced by the nature of the cell extract, which contains various factors on the chromosomal DNA. Some of the Escherichia coli cell extract factors are essential, despite their negative effects on protein synthesis, because they are required during the cell growth and/or extract preparation stage. In this study, modified E. coli strains were generated by inserting a streptavidin binding peptide (SBP) tag sequence at the 3' termini of the genes encoding polynucleotide phosphorylase (PNPase) and/or Exodeoxyribonuclease V alpha chain (RecD) on the chromosomal DNA. The SBP-tagged target gene products were specifically removed from the cell extract prepared from modified E. coli cells using SBP affinity resin. The linear DNA-directed cell-free protein synthesis using the treated extract achieved higher productivity, especially when removing both the PNPase and RecD factors. Using this strategy to remove multiple inhibitory factors in a cell extract will be widely applicable to improve cell-free protein synthesis.

• 公益社団法人日本生物工学会の論文

著者

• Kigawa Takanori

  Nmr Pipeline Methodology Research Team Riken Systems And Structural Biology Center:department Of Com

• Matsuda Natsuko

  Nmr Pipeline Methodology Research Team Riken Systems And Structural Biology Center

• Seki Eiko

  NMR Pipeline Methodology Research Team, RIKEN Systems and Structural Biology Center

• Seki Eiko

  Nmr Pipeline Methodology Research Team Riken Systems And Structural Biology Center:department Of Com

関連論文

• Multiple inhibitory factor removal from an Escherichia coli cell extract improves cell-free protein synthesis(ENZYMOLOGY, PROTEIN ENGINEERING, AND ENZYME TECHNOLOGY)

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