Construction Of A DNA Vaccine PcDNA3.1+/flk-1_nl-4 Against Tumor Angiogenesis

Objective: To develop a novel DNA vaccine pcDNA3.1+/flk-l(n1-4) against tumorangiogenesis.Methods: Extracting total RNA from the tissue of BALB/c mouse embryo andamplifying fik-l(n1-4) cDNA coding 416 amino acids by reverse transcriptase-polymerase chain reaction(RT-PCR). After confirmed by DNA sequence analysis,flk-l(n1-4) cDNA was inserted into eucaryotic expression vector pcDNA3.1 +containing CMV promoter. The recombinant plasmids pcDNA3.1+/flk-l(n1-4) wasconstructed and transfected into eucaryotic expression system COS-7 cells byLipofectamine. Then detecting protein expression of flk-l(n1-4) by Western blot.Results: Extracellular loop 1-4 of flk-1 was cloned and the recombinant plasmidpcDNA3.1+/flk-l(n1-4) was successfully constructed. Both has been proved by DNAsequencing and comparing with data in Gene Bank. Protein expression of flk-l(n1-4)was detected in eucaryotic expression system COS-7 cells.Conclusions : We successfully constructed the DNA vaccine against tumorangiogenesis which can be used to the further study on animal experiment and clinicalapplication.