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Objective: To construct survivin 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-SD 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 expression vector was constructed successfully and got GBC-SD cells which are stable expression shRNA.
Objective: To construct survivin 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 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 expression vector was constructed successfully and got GBC-SD cells which are stable expressi on shRNA.