Construction Of A Nucleic Acid Vaccine PcDNA3.1(+)-flk1-domain1-7 Against Tumor Angiogenesis

Objective To obtain the recombinant plasmid of extracellular ligand binding domains of BALB/c mouse vascular endothelial growth factor receptor -2(VEGFR - 2 /flk -1) and to identify its expressing of protein and to develop a nucleic acid vaccine against extracellular domain of VEGFR-2. Methods Total RNA was isolated from mouse embryo and was subjected to RT - PCR for the amplification of the encoding sequence. The amplified fragment was inserted into eucaryotic expression vector pcDNA3.1(+).Then the recombinant plasmid was transfected into the COS7 cell line and identified its expressing of protein by Western blot analysis. Results (1) We successfully got the extracellular domain of VEGFR-2 which have been proved by DNA sequencing and comparing with data in GenBank. (2) We successfully constructed the recombinant plasmid pcDNA3.1 (+)-flkl-domain1-7 which can express specific protein in eucaryotic expression system and to be a nucleic acid vaccine against tumor angiogenesis. Conclusion We successfully constructed the nucleic acid vaccine pcDNA3. 1(+)-flkl-domainl-7 which will be used to the study of anti-tumor angiogenesis.