PROTEASE INHIBITOR GENE, MICROORGANISM CONTAINING THE GENE AND PRODUCTION OF PROTEASE INHIBITOR USING THE MICROORGANISM

摘要

PURPOSE:To obtain a large amount of the subject gene in high efficiency by culturing a transformant containing a specific DNA fragment. CONSTITUTION:A chromosome DNA fragment originated from Bacillus brevis HPD31 strain (FERM BP-1087) is linked to an enzyme-treated product of vector pUC118 and the linked product is selected by an antibody reaction process to obtain a plasmid pYAS01 (A). The component A is subjected to restriction enzyme treatment and subcloned and the obtained pYAS04 is introduced into an E.coli to obtain E.coli pYAS04 (B) capable of producing a protease inhibitor (PIH). The base sequence of a fragment BamHI-Pst1 of the component B having a size of 1517bp is determined to obtain a DNA sequence (C), etc., corresponding to the amino acid sequence of PIH starting at the 832nd position (TTC: phenylalanine) and ending at the final 1444th position (GAA: glutamic acid). A protease inhibitor can be produced by culturing Bacillus brevis HPD31-S5/pYS 01 transformed with a plasmid originated from the component B.