|Journal of Biological Chemistry (2006) 281(49):37559-65|
|New York Consortium on Membrane Protein StructureNortheast Structural Genomics Consortium|
(click to unfold)
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.
|microbiology metabolism isolation & purification biosynthesis genetics |
|Genes, Bacterial Recombinant Proteins Escherichia coli Genetic Vectors Endoribonucleases DNA-Binding Proteins Plasmids Selenomethionine Nitrogen Isotopes p-Fluorophenylalanine Escherichia coli Proteins Gene Expression Bioreactors |
|29 (Last update: 09/16/2017 11:47:35am)|
|J Biol Chem. 2006 Dec 8;281(49):37559-65. Epub 2006 Oct 4.|