Document Detail


Multiple inhibitory factor removal from an Escherichia coli cell extract improves cell-free protein synthesis.
MedLine Citation:
PMID:  19577188     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Eiko Seki; Natsuko Matsuda; Takanori Kigawa
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of bioscience and bioengineering     Volume:  108     ISSN:  1347-4421     ISO Abbreviation:  J. Biosci. Bioeng.     Publication Date:  2009 Jul 
Date Detail:
Created Date:  2009-07-06     Completed Date:  2009-11-10     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  100888800     Medline TA:  J Biosci Bioeng     Country:  Japan    
Other Details:
Languages:  eng     Pagination:  30-5     Citation Subset:  IM    
Affiliation:
NMR Pipeline Methodology Research Team, RIKEN Systems and Structural Biology Center, Tsurumi-ku, Yokohama 230-0045, Japan.
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MeSH Terms
Descriptor/Qualifier:
Affinity Labels
Bacterial Proteins / biosynthesis*,  isolation & purification
Base Sequence
Cell-Free System
DNA Primers
Escherichia coli / cytology,  genetics,  metabolism*
Chemical
Reg. No./Substance:
0/Affinity Labels; 0/Bacterial Proteins; 0/DNA Primers

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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