MASS EXPRESSION SYSTEM OF MODIFIED SUBSTANCE OF CTOCHROME P450 2C19 IN ESCHERICHIA COLI

摘要

PROBLEM TO BE SOLVED: To mass-produce the subject modified substance, derived from the human liver, having enzymic activities equal to those of a natural cytochrome enzyme P450 2C19 and useful for predicting the presence or absence of medicinal interaction by culturing Escherichia coli having a specific DNA molecule transduced thereinto. ;SOLUTION: This modified resistance of cytochrome enzyme P450 2C19 is obtained by culturing Escherichia coli transformed with an Escherichia coli plasmid containing a DNA molecule containing a DNA sequence ranging from the 1st to the 1416th positions of the formula, solubilizing the resultant bacterial cell and collecting the substance from a membranous fraction and a soluble fraction. The modified substance has the amino acid sequence of the cytochrome enzyme P450 2C19 derived from the human liver in which the 2nd position is substituted by alanine and 18 amino acid residues from the 3rd to the 20th positions are deleted. Thereby, a cDNA of the natural type cytochrome enzyme P450 2C19 can be modified into the DNA sequence by a site-specific mutagenesis according to a polymerase chain reaction(PCR) and a mutational transduction method by insertion of a synthetic oligonucleotide molecule.;COPYRIGHT: (C)1998,JPO