Expression of dengue virus type IIpremembrane and envelope proteins in baculovirus expression system

Objective: Co-expression of dengue virus premembrane and envelope(prM/E) proteins is required for production of virus-like-particles, and the dengue virus-like-particle has become one of the most important aspects in dengue virus vaccine research. Methods: To establish the baculovirus expression system that expresses prM/E proteins, the prM/E gene was obtained by PCR amplification from the plasmid containing the prM/E gene of dengue virustype Ⅱ. The PCR product was cloned into the Xho Ⅰ/ Nhe Ⅰ restriction site of pFastBac Dual vector to establish the transfer vector pFBD-prM/E. Then the pFBD-prM/E was transformed into the competent DH10 Bac containing Bacmid and Helper vector, which established the shuttle plasmid rBacmid-prM/E. The latter was transfected into Sf9 cells, and the recombinant baculovirus was obtained. Results: The expression of prM/E proteins was then confirmed by indirect immunofluorescence assay. Conclusion: The baculovirus expression system expressing prM/E will be useful for further functional studies of prM and E proteins, and development of dengue virus-like-particle vaccine.