Construction of vector TAT-SOX2 and expression of fusion protein

VernacularTitle:Tat融合蛋白表达载体TAT-SOX2的克隆化及蛋白表达研究
Author: Jianjun WANG ¹ ; Zhihong WAN ; Ping ZHAO ; Xueyuan JIN ; Guoming XIE ; Shaojie XIN
Author Information

1. 解放军医学院
Keywords: Protein SOX2; Stem cells; Protein transprot
From: Chinese Journal of Experimental and Clinical Virology 2014;28(5):352-354
CountryChina
Language:Chinese
Abstract: Objective To construct recombinant expression vector TAT-SOX2 and express the fusion protein in E.coli BL21.Methods SOX2 fragment were amplified by PCR.The product was digested with restriction enzymes then was inserted into PET-28b-TAT-V2 which contains protein transduction domain of TAT.The recombinant vector was transformed into E.coli BL and induced with IPTG.The expressed fusion protein was analyzed by using SDS-PAGE method.The highly expressed product was purified by affinity chromatography.Western blotting was used to identify the specificity of the fusion protein.The immunofluorescence was used to detect the efficiency of fusion protein transduced to HSF cells.Results The recombinant expression vector TAT-SOX2 was correctly constructed and fusion protein TAT-SOX2 was successfully expressed in prokaryotic cells.Western blotting showed the specificity of the fusion protein.The immunofluorescence prompt fusion protein was transduced into HSF cells quickly.Conclusion This study provides the material basis for the further induced pluripotent stem cells by protein transduction.