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Multiple inhibitory factor removal from an Escherichia coli cell extract improves cell-free protein synthesis

Paper ID Volume ID Publish Year Pages File Format Full-Text
21747 43238 2009 6 PDF Available
Title
Multiple inhibitory factor removal from an Escherichia coli cell extract improves cell-free protein synthesis
Abstract

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.

Keywords
Cell-free protein synthesis; RecBCD; PNPase; Red/ET recombination; Streptavidin binding peptide tag
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Multiple inhibitory factor removal from an Escherichia coli cell extract improves cell-free protein synthesis
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 108, Issue 1, July 2009, Pages 30–35
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us