Construction and initial application of recombinant adenoviral expressed vector carrying EBV immediately early gene BZLF1
Objective To construct recombinant adenoviml express vector carrying EBV immediately early gene BZLF1 and to investigate its function of inducing latent EBV into lytic cycle in vitro. Methods The recombinant adenovirus vector carrying EBV immediately early gene BZLF1 was constructed with AdEasy system and then transfected 293 cells to create recombinant adenovirus vAd-BZ. EBV positive NEC cells were infected with recombinant adenoviruses. The expression of target gene and its effects on EBV positive cells were tested by RT-PCR, Western blot, FACS and MTT in vitro. Results The recombinant adenovirus vector was constructed successfully. The recombinant adenovirus vAd-BZ, which has stable infectivity, was packed in 293 cells. BZLF1 can be detected in EBV positive cells NEC infected by recombinant adenovirus and EBV was induced to enter lytic form after infection. Conclusion The recombinant adenoviruses vAd-BZ can effectly infect EBV positive NEC cells, induce a switch of EBV from latency to lyric cycle and kill EBV positive tumor cells specifically, which is potential for gene therapy of EBV associated tumor.