MODIFIED HUMAN TUMOR MECROSIS FACTOR

摘要

PURPOSE:To readily put remedy of antitumor, etc., to practical use in high activity by substituting amino acid at carboxyl end of human tumor necrosis factor of wild-type with other amino acid. CONSTITUTION:SalI broken site at carboxyl end of structural gene in human tumor necrosis factor (hTNF) of wild-type (natural type) and BalI site at 40 bp upstream from said broken site are utilized and leucine in amino acid is substituted with phenylalanine, then DNA coding carboxyl end mutant is chemically synthesized to obtain DNA fragment (A) expressed by the formula. Next, A-component is itroduced into plasmid pUC19 (B) and vector (C) as plasmid pATNF-CF is obtained. Then, C-component is introduced into Escherichia coli and Escherichia coli K12C600(m-R-)/pATNF-CF (D) as transformant is obtained. Thus, D-transformant is cultured in desired medium and resultant cultured matter is purified by centrifugal separation or chromatography, etc., to produce modified human tumor necrosis factor.