RECOMBINANT PLASMID DNA PTRCTE-OPH AND PRODUCER OF ENZYME ORGANOPHOSPHATE HYDROLASE Russian patent published in 2004 - IPC

FIELD: biotechnology, genetic engineering, biochemistry.

SUBSTANCE: invention represents the constructed in vitro recombinant plasmid DNA comprising for the processed form of enzyme organophosphate hydrolase (OPH), Escherichia coli trc-promoter and synthetic site as an enhancer of translation providing biosynthesis of enzyme OPH, and the strain Escherichia coli as a producer of this protein also. Recombinant enzyme can be used for producing OPH-base preparations designated for decomposition of organophosphorus compounds and for their analytical determination also. Invention solves the problem for the development of genetic-engineering construction and strain of cells that provide the preparing significant amounts of active soluble form of organophosphate hydrolase in cell cytoplasm by inducible synthesis of enzyme. This problem is solved by construction of recombinant DNA pTrcTE-OPH encoding inducible synthesis of OPH and the strain of Escherichia coli DH5α/pTrcTE-OPH providing synthesis of this protein with the expression level allowing to prepare up to 12 mg of purified protein from 1 g of wetted biomass. High inducible level of synthesis of the end polypeptide is provided by the fact that the proposed plasmid pTrcTE-OPH differs from the known one by the presence of E. coli trc-promoter and synthetic enhancer of translation for bacteriophage T7 gene 10.

EFFECT: valuable biological properties of DNA.

3 cl, 2 tbl, 4 dwg, 3 ex