RAT RENAL P450C24-PRODUCING BACTERIAL STRAIN

摘要

PURPOSE:To obtain a expression plasmid for producing P450C24 used in synthesis of a vitamin D3 derivative by bonding a rat renal P450C24 gene to the downstream of Escherichia coli trp-lac (tac) promotor and expressing rat renal P450C24 in Escherichia coli bacterium. CONSTITUTION:The objective rat renal P450C24 WO is produced in large amounts by extracting a genome DNA from a rat renal mitochondria according to ordinary method, selecting a gene coding a rat renal mitochondria P450C24 from the extract, bonding the gene to the downstream of Escherichia coli trp-lac (tac) promotor, assembling a expression plasmid pKC24R expressed by the formula and capable of expressing rat renal P450C24 in Escherichia coli bacterium, inserting the expression plasmid into Escherichia coli JM109 strain to transform the strain and then culturing the transformant in a culture medium to manifest the expression plasmid. Furthermore, the P450C24 is conjugated with a bovine adrenal adrenodoxin and a bovine adrenal NADPH-adrenodoxin reductase to provide a vitamin D3 derivative hydroxylated in 24 position.