CYTOCHROME P-450HFLA PROTEIN, DNA SEQUENCE THEREOF AND PRODUCTION THEREOF

摘要

PURPOSE:To make it possible to obtain the title protein in large quantities without containing a human-derived other protein by culturing a recombinant host cell in a culture medium. CONSTITUTION:A human fetal lever is purified by homogenizing in a buffer to afford a cytochrome P-450HFLa protein (A) having about 51000 molecular weight. Then a rabbit, etc., is immunized with the ingredient A to give an anti-P-450HFLa antibody (B). On the other hand, a fetal lever is treated with guanidinethiocyanate to extract total RNA, which is then chromatographed to afford mRNA (C). Then the ingredient C is translated to a protein and subjected to screening with the ingredient B to provide the desired mRNA fraction (D). Then the ingredient D is synthesized to a single-stranded cDNA, which is then treated with ligase to give a double-stranded cDNA (E). Then a recombinant DNA obtained by adding a linker to the ingredient E and introducing into a plasmid is introduced into escherichia coli and the resultant transformed strain is cultured in a culture medium and DNA sequence such as DNA expressed by formula I is expressed to produce cytochrome P-450HFLa protein containing an amino acid sequence expressed by formula II.