Comparison of two techniques for expression and purification of glycogen synthase kinase 3β.
- Author:
Shao-fei XU
1
;
Jie XU
;
Ming-tao LI
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Baculoviridae; metabolism; Escherichia coli; metabolism; Genetic Vectors; Glycogen Synthase Kinase 3; biosynthesis; isolation & purification; Insecta; cytology
- From: Journal of Southern Medical University 2011;31(3):397-402
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo establish a method for the expression of glycogen synthase kinase 3β with high purity and biological activity.
METHODSE.coli expression system and baculovirus-insect cell expression system were used to produce the kinase, followed by purification using His-tag and GST-tag and determination of its purity and activity by SDS-PAGE and kinase reaction, respectively.
RESULTSGlycogen synthase kinase 3β produced from E.coli represented 54% of the total bacterial protein, as compared with 96% of the total protein from the insect cell system .Glycogen synthase kinase 3β produced from insect cell exhibited an one-fold higher biological activity than the protein obtained from E.coli.
CONCLUSIONSCompared with the protein from E.coli system, glycogen synthase kinase 3β from the insect cell expression system is endowed with a higher purity and bioactivity.