Construction of Eukaryotic Expression Vector for EGFP Gene Modulated by Survivin Promoter and Its Specific Expression in Human Lung Cancer Cells

Objective To construct a eukaryotic expression vector for enhanced green fluorescent protein(EGFP)gene modulated by survivin promoter and identify its specific expression in human lung cancer A549 cells. Methods Survivin promoter was amplified by PCR using the genomic DNA of A549 cells as a template,with which the CMV promoter in pEGFP-C1 vector was substituted. The constructed recombinant plasmid pEGFP-C1 / Surp was transfected to A549 and MRC-5 cells,and the expression of EGFP was observed by fluorescent microscopy. Results Restriction analysis and sequencing proved that recombinant plasmid pEGFP-C1 / Surp was constructed correctly. Strong green fluorescence was observed by microscopy in the A549 cells transfected with recombinant plasmid pEGFP-C1 / Surp. However,no green fluorescence was observed in transfected MRC-5 cells. Conclusion A eukaryotic expression vector for EGFP gene modulated by survivin promoter was successfully constructed,and survivin showed strong tumor-specific promoting activity. It laid a foundation of further development of vector for gene therapy targeting tumors.