APin 재조합 단백질의 합성과 적용

Abstract

Apin protein, calcifying epithelial odontogenic (pindborg) tumors (CEOTs)-associated amyloid, were isolated from CEOTs, and has similar nucleotide sequences to OD314. It was reported that APin was highly expressed during ameloblast maturation and mineralization. APin protein was also efficiently secreted from cultured ameloblasts. However, little is known about the functional study of APin using recombinant protein during amelogeneis. In this study, in order to produce the APin recombinat protein, the APin-expression construct were made and expressed its protein in various host cell and temperature conditions for the utilization of further enamel functional studies. The results were as follows: 1. APin protein was strongly expressed in the induction system using pRSET-OD314 construct. 2. When the JM109 was used as a expression host, APin protein was strongly expressed in the induction system using pHCEIIBNd-OD314 construct. 3. The APin protein was recognized at a molecular weight of 22 kDa in Western blots. 4. Almost of the expressed APin protein was soluble. These results suggest that considerable amount of APin recombinat protein was produced and it could be used for further amelogenesis research effectively.