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Expression of Recombinant Protein of Human PAK1 and Its Location in Cytoplasm of BGC-823 Cells
Gui-Ling Wang, Ying Zhou, Da-Lin Zhao1, Jia-Bin Li, Fu-Rong Liu, Feng Li*
Key Laboratory o f Cell Biology, Ministry of Public Health, China Medical University, Shenyang 110001, China; 1Department of Physiology, Shenyang Institute of Athletics, Shenyang 110001, China
Abstract: To detect p21-activated kinase1 (PAK1) expression in prokaryocyte and its location in transfected eukaryotic cells， we obtained PAK1 gene by RT-PCR and cloned the coding sequences of PAK1 into the procaryotic expression vector pGEX-5X-1.The expressed proteins were induced by IPTG and identified by SDS-PAGE ， Western blot and kinase assay in vitro. At the same time， GFP-tagged eukaryotic expression vector pEGFP/PAK1 was constructed and then transfected into human gastric cancer cell line BGC-823 with liposome， and the expression of GFP was observed by confocal laser scanning microscopy. pGEX-5X-1/PAK1 could express GST-PAK1 fusion protein with 94kDa. Under microscopy， the expression of GFP-tagged PAK1 was observed in cytoplasm of BGC-823 cells transfected with pEGFP/PAK1. This study provides the basis for research on biological features of PAK1.
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