Construction of recombinant adenoviras expressing RSV F gene

Abstract

Objective To construct a recombinant adenovirus contained respiratory syncytial virus F gene fragment. Methods The F gene fragment was amplified by RT-PCR. Then the recombinant shuttle plasmid pShuttle-CMV/F was constructed. The iinearized pShuttle-CMV/F was transformed into BJ5183-AD-1 electroporation competent cells which contained plasmid pAdEasy-1. The obtained homologous recom-binant plasmid was named pAdEasy/F, and transfected into 293 cells. Then the recombinant adenovirus rAd/F was obtained, and it was tested by electron microscope, RT-PCR, Western blot and immuno-fluores-cence assay(IFA). The vires titer and genetic stability were also studied initially. Results The recombi-nant adenovirus rAd/F was constructed, which was visualized as typical adenovirus morphology under elec-tron microscope. The transcription and expression of RSV F gene fragment integrated in the rAd/F were con-finned by RT-PCR, Western blot and IFA. Conclusion The recombinant adenovirus rAd/F contained RSV F gene fragment is successfully constructed and lay an important foundation for the further vaccine study. Key words: Respiratory syncytial virus; Fusion protein; Adenovirus vector