Development of a stable insect cell line constitutively expressing rotavirus VP2

An insect High-Five cell line was generated constitutively and stably expressing the VP2 protein of rotavirus RF strain leading to the formation of core-like particles. The integration of the VP2 gene was confirmed by PCR, Real-time PCR and Southern blot, and the protein expression was confirmed by Western blot and immunofluorescence microscopy. Integrity and self assembly of VP2 as core-like particles was demonstrated by electron microscopy. The High-Five cell system stably expressing rotavirus core-like particles can be applied to the study of viral protein structure and functions; it may be useful for vaccine development, gene therapy and drug delivery.

► VP2 gene of rotavirus cloned in the dual-host expression vector pBiEx-3.
► The expression of VP2 from recombinant pBiEx-3 confirmed in E. coli BL-21 (DE3).
► High-Five insect cells transfected with VP2-recombinant pBiEx-3.
► PCR, Real-time PCR, Southern blot, Western blot and immunofluorescence microscopy confirmed presence of VP2.
► Self-assembly of VP2 proteins observed by electron microscopy in stably transfected High-Five insect cell clones.