Patent ID: 8268590

Claim:
A method for preparing non-activated tissue-regeneration polypeptide (TRP), comprising the steps of: (a) culturing transformed bacteria to express [PTD-FAD-TRD] a polypeptide in a culture broth, wherein the transformed bacteria are bacteria transformed with a recombinant bacterial expression vector comprising a furin activation domain (FAD)-encoding base sequence upstream of the 5′ region of tissue regeneration domain (TRD)-encoding DNA, a protein transduction domain (PTD) base sequence, a base sequence for tagging, and at least four histidine-encoding base sequences for separation and purification, wherein the PTD is a TAT sequence selected from the group consisting of SEQ ID NOs: 27 and 43, the TRD is an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 3 and 6 and the FAD is an amino acid sequence selected from the group consisting of SEQ ID NOs: 14 and 19; and (b) centrifuging the culture broth; (c) removing the secondary or tertiary structure of the polypeptide or converting the secondary or tertiary structure to primary linear structure by the addition of a urea solution into the supernatant and cell pellet to generate a urea, supernatant, cell pellet mixture; and (d) separating from the mixture and purifying a non-activated polypeptide.