Cloning and Expression of VP2 Gene of the Bovine Rotavirus RF in a Mammalian Cell Line

Rotaviruses are recognized as the most important cause of sever diarrhea illness of infants and children world-wide. These viruses consist of triple-layered protein capsid The inner layer is composed of VP2. In this study, the expression of VP2 gene of the bovine rotavirus RF has been evaluated in human epithelial lung cells (A549) under CMV promoter. First, the rotavirus gene was cloned in pcDNA3 plasmid and then recombinant plasmid transferred to a human lung epithelial cell line by a carrier namely dendrosome. Western blotting analysis has revealed expression of the gene in the cells after 48-72 hrs post infection. The expression of VP2 gene was confirmed by observation of core like particles (CLPs) with electron microscopy and immunocytochemistry by using gold conjugated protein A. The results indicated that the used eukaryotic cells in this study can serve as a suitable host for expression of VP2 gene; since the cells expressed the proteins correctly and made the VP2 protein to be self-assembled and form native structure.