Development the platform of IMPACT expression system

• Main Authors: Ming Han Wu, 吳明翰
• Other Authors: Chao-Lin Lin
• Format: Others
• Language: zh-TW
• Published: 2008
• Online Access: http://ndltd.ncl.edu.tw/handle/14017309692357995578

碩士 === 明志科技大學 === 生化工程研究所 === 96 === Easy manipulation, efficient production and lower cost have made E.coli frequently utilized as the platform for protein expression. In this study, we used pRSET, pTriex and pTwin as our expression vector. pRSET is a high copy plasmid. pTriex vector could be expressed in E.coli, Baculovirus, and mammalian cells. Furthermore, pTwin vector could express target proteins without adding any redundant amino acids. Band 3 protein is an anion channel protein located on the RBC membrane and identified as the autoantigen of patients with SLE. The binding of autoantibodies against Band 3 protein would result in autoimmune haemolytic anemia. The Band3 protein sequence 861 to 874 is thought to be processed and presented by antigen present cells to activated naïve T cells. In this study, we optimized the expression of the proteins including Band 3 peptide (Band 3 217t2916), Band3 peptide (1913t2916), Band 3 protein epitope homologues CESP9 (CESP 1320666t132062) and CESP30 (CESP 1320643t1320719) from helicobacter pylori by pRSET, pTriex and pTwin in E.Coli expression system.