Construction of a Recombinant Bacmid DNA to Express Influenza Virus Matrix Protein1 (M1) in Insect Cell Line

Virus-like particles (VLPs) have been suggested to be a promising recombinant vaccine approach. Several studies have reported that the influenza VLPs produced in insect cells is an effective vaccine candidate. Due to crucial role of matrix M1 protein in assembly and budding of Influenza particles, in all VLPs structures, M1 protein have been considered as a main component. M1 open reading frame (759 bp) from human influenza virus A /New Caledonia 20/1999/ (H1N1) was amplified by RT-PCR. The amplicone was cloned into pFastBac1 donor plasmid through KpnI/XhoI sites. After verification of clone by restriction analysis, it was subjected to automated sequencing. The M1 recombinant bacmid was subsequently generated and verified by PCR using M1 specific and M13 universal primers. results showed that a recombinant baculovirus containing correct and inframe sequence of Influenza M1 gene under control of polyhedrin promoter has been constructed. The above-mentioned M1 recombinant baculovirus can be used with other individual recombinant baculoviruses expressing HA and NA genes to produce influenza VLPs in insect cell line.