RECOMBINANT CPP-32 GENE, EXPRESSION VECTOR CONTAINING THE SAME, AND METHOD FOR MANUFACTURING ACTIVE CPP-32 PROTEIN IN E.COLI BY USING THE SAME

摘要

PURPOSE: Provided are recombinant CPP(cysteine putative protease)-32 gene, an expression vector containing the same, and a method for manufacturing active CPP-32 protein in E.coli by using the same, thereby simultaneously expressing p12 and p17 subunits and producing active CPP-32 in E.coli massively. CONSTITUTION: A vector pRGT7His7 CPP-32/p12-p17 contains T7 promotor, p12 coding gene, a nucleotide sequence of SEQ ID NO:5 encoding isoleucine-aspartic acid-glutamic acid-phenylalanine, and p17 coding gene. It expresses recombinant CPP-32 protein by bicistronic expression method. A transformed E.coli BL21(DE3)/pRGT7His7 CPP-32/p12-p17(KCTC-0479BP) is obtained by transformation with the above vector. A method for manufacturing the recombinant CPP-32 protein comprises culturing the transformed E.coli, followed by cytolysis, centrifuging the crude lysate to remove insoluble protein, and performing chromatography.