Please use this identifier to cite or link to this item:
|標題:||Production of recombinant EGFP via surface display of ice nucleation protein and self-cleavage intein||作者:||Wu, J.Y.
|Project:||Biochemical Engineering Journal||期刊/報告no：:||Biochemical Engineering Journal, Volume 54, Issue 3, Page(s) 158-163.||摘要:||
In this study, a novel recombinant protein production system was developed by constructing a protein chimera with a self-cleavage segment along with a cell surface display segment to finally produce a model protein entity (i.e., enhanced green fluorescent protein, EGFP). In the plasmid construction, the EGFP gene was fused to genes of a self-cleaving intein (INT) and an ice nucleation protein (INP) which can anchor on the cell membrane. In the cultivation of recombinant Escherichia coil, the cells expressed high green florescence after the addition of isopropyl-beta-D-thiogalactopyranoside (IPTG) as the inducer. By simply holding the cell pellets in Tris-HCl (pH 10.0) buffer at room temperature, EGFP was solubilized from the INP-INT segment embedded on the cell surface via intein's self-cleavage function. The EGFP concentration of 273 mg/L and recovery of 88% were obtained at day 5, whereas the best EGFP productivity of 187 mg/L/d was obtained at day 1. The EGFP can be harvested only via centrifugation, and no cell disruption process is required. This simplified approach is expected to be applicable for obtaining recombinant functional proteins for academic and industrial use. (C) 2011 Elsevier B.V. All rights reserved.
|Appears in Collections:||分子生物學研究所|
Show full item record
TAIR Related Article
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.