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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
3
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pubmed:dateCreated |
1996-10-24
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pubmed:abstractText |
We modified a cell-free coupled transcription/translation system from Escherichia coli with the T7 phage RNA polymerase, and achieved a productivity as high as 0.4 mg protein/ml reaction mixture. First, we found that the optimal concentrations of phosphoenolpyruvate and poly(ethylene glycol) are interdependent; higher concentrations of the former should be used at higher concentrations of the latter. Second, the use of a condensed 30000 x g cell extract, in place of the conventional one, significantly increased the initial rate of protein synthesis. This phenomenon was demonstrated to be due to a reason other than elimination of inhibitory molecule(s) from the extract. For this system with the condensed extract, the phosphoenolpyruvate and poly(ethylene glycol) concentrations were again co-optimized, resulting in production of chloramphenicol acetyltransferase at a productivity of 0.3 mg/ml. Finally, the productivity was further increased up to 0.4 mg/ml, by supplementation of the pool of amino acids. This improved cell-free protein synthesis system is superior in productivity to any other cell-free systems reported so far, including the continuous-flow cell-free system.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Chloramphenicol O-Acetyltransferase,
http://linkedlifedata.com/resource/pubmed/chemical/DNA-Directed RNA Polymerases,
http://linkedlifedata.com/resource/pubmed/chemical/Magnesium,
http://linkedlifedata.com/resource/pubmed/chemical/Phosphoenolpyruvate,
http://linkedlifedata.com/resource/pubmed/chemical/Polyethylene Glycols,
http://linkedlifedata.com/resource/pubmed/chemical/Viral Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/bacteriophage T7 RNA polymerase
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pubmed:status |
MEDLINE
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pubmed:month |
Aug
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pubmed:issn |
0014-2956
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:day |
1
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pubmed:volume |
239
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
881-6
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pubmed:dateRevised |
2007-7-23
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pubmed:meshHeading |
pubmed-meshheading:8774739-Cell-Free System,
pubmed-meshheading:8774739-Chloramphenicol O-Acetyltransferase,
pubmed-meshheading:8774739-DNA-Directed RNA Polymerases,
pubmed-meshheading:8774739-Escherichia coli,
pubmed-meshheading:8774739-Magnesium,
pubmed-meshheading:8774739-Phosphoenolpyruvate,
pubmed-meshheading:8774739-Polyethylene Glycols,
pubmed-meshheading:8774739-Protein Biosynthesis,
pubmed-meshheading:8774739-Transcription, Genetic,
pubmed-meshheading:8774739-Viral Proteins
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pubmed:year |
1996
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pubmed:articleTitle |
A highly efficient cell-free protein synthesis system from Escherichia coli.
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pubmed:affiliation |
Interdisciplinary Program for Biochemical Engineering and Technology, College of Engineering, Seoul National University, Korea.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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