PRODUCTION OF THROMBIN-INHIBITING SUBSTANCE

摘要

PURPOSE:To enable mass-production of hirudin having anti-thrombin activity by using a DNA sequence produced by bonding a hirodin-coding DNA fragment to a site immediately after a specific region of a neutral protease gene of a bacterium of genus Bacillus. CONSTITUTION:A hirudin-secreting plasmid is produced by bonding a hirudin- coding DNA fragment to a site immediately after the promoter, a ribosome- bonding site and a secretion signal coding region of a neutral protease gene of a bacterium of genus Bacillus (preferably Bacillus amyloliquefaciens) and bonding the obtained DNA sequence (preferably the sequence of formula) to a vector DNA. A host microorganism (preferably Bacillus subtilis) is transformed with the produced secretion plasmid and the transformant is cultured. Hirudin is separated from the supernatant liquid of the cultured product.