|Journal of Biological Chemistry (2006) 281(49):37559-65|
|New York Consortium on Membrane Protein StructureNortheast Structural Genomics Consortium|
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We developed a new bacterial expression system that utilizes a combination of attributes (low temperature, induction of an mRNA-specific endoribonuclease causing host cell growth arrest, and culture condensation) to facilitate stable, high level protein expression, almost 30% of total cellular protein, without background protein synthesis. ...
With the use of an optimized vector, exponentially growing cultures could be condensed 40-fold without affecting protein yields, which lowered sample labeling costs to a few percent of the cost of a typical labeling experiment. Because the host cells were completely growth-arrested, toxic amino acids such as selenomethionine and fluorophenylalanine were efficiently incorporated into recombinant proteins in the absence of cytotoxicity. Therefore, this expression system using Escherichia coli as a bioreactor is especially well suited to structural genomics, large-scale protein expressions, and the production of cytotoxic proteins.
|metabolism genetics isolation & purification biosynthesis microbiology |
|Escherichia coli Escherichia coli Proteins Recombinant Proteins Genes, Bacterial DNA-Binding Proteins Selenomethionine Nitrogen Isotopes Gene Expression Genetic Vectors Endoribonucleases Plasmids Bioreactors p-Fluorophenylalanine |
|39 (Last update: 03/16/2019 7:20:09pm)|
|J Biol Chem. 2006 Dec 8;281(49):37559-65. Epub 2006 Oct 4.|