Synthesis of DNA Encoding Botulinum Neurotoxin Receptor syt II and Its Fusion Expression in E.coli
- VernacularTitle:肉毒毒素蛋白受体sytIIN端片段基因的合成及其在大肠杆菌中的融合表达
- Author:
Jing SHI
;
Hui WANG
;
Shi-Zhong BAO
;
Xiao-Jun HOU
;
Jun YIN
;
- Publication Type:Journal Article
- Keywords:
Botulinum neurotoxin Receptor MBP fusion protein Prokaryotic expression
- From:
China Biotechnology
2006;0(07):-
- CountryChina
- Language:Chinese
-
Abstract:
In order to construct a prokaryotic expression vector of human receptor syt II N-fragment and to express recombinant MBP-Syt fusion protein in E.coli and to purify and identify its activity. According to codon preference of E.coli, a DNA fragment encoding human syt II N-fragment was synthesized, and then cloned into prokaryotic vector pMAL-c2x for sequencing. Then the recombinant plasmid pMAL-Syt was introduced into E.coli ER2566 by transformation for expression and the obtained engineered bacteria were induced by IPTG. The fusion protein was purified by amylose resin affinity chromatography and identified by SDS-PAGE and Western blot. The binding activity of the protein was determined by ELISA. It is concluded that MBP-Syt protein is of good binding activity.
