Construction and Expression in Vitro of Recombinant Chicken Survivin Adenoviral Vector

Total RNA was isolated from chicken embryos and subjected to RT-PCR for amplification of chicken survivin using primers according to it's cDNA sequence in GenBank.The amplified product was inserted into T-A clone vector and then subcloned into adenoviral shuttle vector and adenoviral backbone vector.The recombinant adenoviral plasmid was transfected into 293E4pIX cells to generate recombinant chicken survivin adenoviral vector.The sequence of chicken survivin was confirmed by sequencing to be identical with that in GenBank and the recombinant adenoviral plasmids were screened by restriction enzyme digestion.Further,PCR and Western blot analysis was used to confirm the expression of survivin in the transfected cells.The recombinant chicken survivin adenoviral vector was constructed successfully and could be further used in tumorimmunotheapy.