Objective： To construct sarvivin shRNA expression vector carting enhanced green fluorescent protein gene, transfect it into GBC-SDH cells via electroporation, and get GBC-SD cells which are stable expressing survivin...Objective： To construct sarvivin shRNA expression vector carting enhanced green fluorescent protein gene, transfect it into GBC-SDH cells via electroporation, and get GBC-SD cells which are stable expressing survivin shRNA. Methods： The siRNA sequence targeting survivin mRNA was synthesized and cloned into pEGFP-H1. The constructed plasmid and pEGFP-H1 were transfected into GBC-SI） cells respectively via liposome, and the transfecting effect was detected with Flow Cytometry. Then the transfected cells were selected with G418. Results： The recombinant plasmid was successfully constructed, named pEGFP-survivin. The gene transfection efficiencies in pEGFP-H1-transfected group and pEGFP-survivin- transfected group were the 80.29% ± 2.71% and 83.85% ±2.34%（P〉0.05）, which was successful to get the cells that are stable expressing shRNA, named GBC-SD/EGFP and GBC-SD/survivin. Conclusion： Survivin shRNA successfully and got GBC-SD cells which are stable expression vector was constructed expression shRNA.展开更多
文摘Objective： To construct sarvivin shRNA expression vector carting enhanced green fluorescent protein gene, transfect it into GBC-SDH cells via electroporation, and get GBC-SD cells which are stable expressing survivin shRNA. Methods： The siRNA sequence targeting survivin mRNA was synthesized and cloned into pEGFP-H1. The constructed plasmid and pEGFP-H1 were transfected into GBC-SI） cells respectively via liposome, and the transfecting effect was detected with Flow Cytometry. Then the transfected cells were selected with G418. Results： The recombinant plasmid was successfully constructed, named pEGFP-survivin. The gene transfection efficiencies in pEGFP-H1-transfected group and pEGFP-survivin- transfected group were the 80.29% ± 2.71% and 83.85% ±2.34%（P〉0.05）, which was successful to get the cells that are stable expressing shRNA, named GBC-SD/EGFP and GBC-SD/survivin. Conclusion： Survivin shRNA successfully and got GBC-SD cells which are stable expression vector was constructed expression shRNA.
