PRODUCTION OF NOVEL PLASMID, MICROBIAL CELL AND ANTITUMOR ACTIVE POLYPEPTIDE

摘要

PURPOSE:To readily and efficiently enable extracellular secretion of a micro organism by transforming a microorganism with a DNA capable of coding a specific peptide and culturing the resultant microorganism in a culture medi um. CONSTITUTION:A DNA fragment obtained by cloning a human antitumor active gene is linked to introduce (A) the firs DNA region containing a DNA sequence (hereinafter referred to as DNA) capable of coding an antitumor active polypep tide (TNFP) expressed by the formula, a promoter DNA derived from a chromosomic DNA (BacDNA) of 170 strains of alkalophilic Bacillus No.170 strain capable of regulating expression of the TNFP and a DNA capable of coding a signal peptide of the BacDNA and (B) the second DNA region containing a DNA, capable of promoting extracellular secretion and derived from a plasmid pMB9 and a promoter DNA capable of regulating expression of the DNA into a plasmid to afford (C) pBXTNF9. Escherichia coli HB101 strain transformed with the component (C) is then aerobically cultured in an LB culture medium, etc., at pH5-8 for 12-48hr to produce the antitumor active polypeptide on the outside of the microbial cells.