A Method for Overproduction of a Protein of a Target Gene in Escherichia coli (Biological Chemistry)

概要

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A simple method for overproduction of a target protein by genetic engineering techniques has been establised. This method involves rearranging the target gene, which contains a ribosome binding sequence for expression, inplurally repeated form, and inserting it in a 3' lower part of promoters. The chloramphenicol acetyltransferase (CAT) structural gene was used to demonstrate the validity of this method. E. coli harboring a CAT expression plasmid, pUS(CAT)_1, which had one inserted CAT gene, was able to produce CAT at the level of only 4% of the total cellular protein according to densitometric scanning on Coomassie-blue-stained SDS-polyacrylamide gel and had the CAT activity of 3.9×10^3 units/mg protein. However, E. coli harboring a CAT expression plasmid, pUS(CAT)_4, which had inserted four directly repeated copies of the CAT gene, could synthesized CAT up to 16% of the total cellular protein and had the CAT activity of 2.8×10^4 units/mg protein. This suggests that this method should be useful for overproducing many important peptides of protein in bacteria.
社団法人日本農芸化学会の論文
1988-09-23