Construction and identification of recombinant fowlpox virus of coexpressing HIV-1_CN gp120 and hIL-6

Objective: To construct the recombinant fowlpox virus (rFPV) coexpressing of HIV-1_CN gp120 and hIL-6. Methods: pUTA-GP-IL6 was constructed by inserting HIV-1 gp120 and hIL-6 gene into the downstream of combined promotor ATI-P7.5 and P7.5 tandem promotor of fowlpox virus expression vector pUTAL. The recombinant plasmid and FPV 282E4 strain were cotransfected chicken embryo fibroblast (CEF) via liposome. The recombinant virus was identified by PCR, indirect immunofluorescent assay (IFA) and Western blot after three times of pressure selection with BUdR, and the mice immunization experiment was performed. Results: 1.4kb fragment could be amplified in the genome of rFPV, and the green fluorescence existed on the cellular surface infected by rFPV. Western blot analysis of expressed products showed that rFPV expressed gp120 and hIL-6 protein. rFPV could elicit the specific humoral immune response. Conclusion: rFPV coexrpessing HIV-1 gp120 and I hIL-6 has been successfully constructed which lays the basis on the preparation of live vector genetic engineering vaccine against HIV-1.