Expression, purification, and bioactivity of GST-fused v-Src from a bacterial expression system  

作　　者：龚兴国 纪静 谢捷 周远 张俊彦 钟文涛 

机构地区：[1]Institute of Biomacromolecule and Enzyme Engineering, School of Life Sciences Zhejiang University,Hangzhou 310027, China

出　　处：《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》2006年第1期13-19,共7页浙江大学学报（英文版）B辑（生物医学与生物技术）

基　　金：Project (No. 03C26213300586) supported by the National Innova-tion Fund for Technology, Ministry of Science and Technology of China

摘　　要：v-Src is a non-receptor protein tyrosine kinase involved in many signal transduction pathways and closely related to the activation and development of cancers. We present here the expression, purification, and bioactivity of a GST (glutathione S-transferase)-fused v-Src from a bacterial expression system. Different culture conditions were examined in an isopropyl β-D-thiogalactopyranoside (IPTG)-regulated expression, and the fused protein was purified using GSH (glutathione) affinity chromatography. ELISA (enzyme-linked immunosorbent assay) was employed to determine the phosphorylation kinase activity of the GST-fused v-Src. This strategy seems to be more promising than the insect cell system or other eukaryotic systems em- ployed in earlier Src expression.v-Src is a non-receptor protein tyrosine kinase involved in many signal transduction pathways and closely related to the activation and development of cancers. We present here the expression, purification, and bioactivity of a GST （glutathione S-transferase）-fused v-Src from a bacterial expression system. Different culture conditions were examined in an isopropyl ,B-D-thiogalactopyranoside （IPTG）-regulated expression, and the fused protein was purified using GSH （glutathione） affinity chromatography. ELISA （enzyme-linked immunosorbent assay） was employed to determine the phosphorylation kinase activity of the GST-fused v-Src. This strategy seems to be more promising than the insect cell system or other eukaryotic systems employed in earlier Src expression.

关 键 词：V-SRC GST-fusion Inclusion body ORTHOGONALIZATION Protein tyrosine kinase 

分 类 号：Q78[生物学—分子生物学]