A construction strategy for a baculovirus-silkworm multigene expression system and its application for coexpression of type I and type II interferons

Liu, Xingjian; Yang, Xin; Mehboob, Arslan; Hu, Xiaoyuan; Yi, Yongzhu; Li, Yinü; Zhang, Zhifang

In recombinant baculovirus, polyhedrin promoter is the preference for foreign gene expression. The polyhedrin gene is replaced by a foreign gene. The saved cost of polyhedrin expression provides for the recombinant protein expression. In reBm-Cαγ, the deletion of p10 gene can also save a certain degree of cost of gene expression, even though the p10 promoter is weaker than the polyhedrin promoter. And these saving costs could be used for foreign gene The antiviral activity of the reBm-Cαγ product was approximately 5-10 times greater than that of the reBm-Cα or reBm-Cγ product. After acid and heat treatment (pH 2.0, 56°C), the antiviral activity of the reBm-Cα product was almost the same as that before the treatments. The antiviral activity of the treated reBm-Cγ product was negligible. The antiviral activity of the treated reBm-Cαγ product was greater than that of chIFN-α. It was approximately 1.7 times greater than that of the reBm-Cα product ily. This multigene baculovirus expression system can be a potential tool in many research fields. The BES is advantageous for antibody expression (Verma, Boleti, & George, 1998) . Previously, heavy-and light-chain genes were inserted in the same gene site (Furuta et al., 2010) . Using the multigene expression strategy in the present study, heavy-and light-chain genes can be introduced into different sites in one recombinant baculovirus, and enhanced expression levels will be achieved. rAAV packaging is another application area of the BES. In preceding studies, rAAV packaging required two or three recombinant baculoviruses, which individually contained cap, rep, and target genes (Aslanidi, Lamb, & Zolotukhin, 2009; Negrete et al., 2007) . By employing our multigene expression strategy, rep, cap, and target genes can be inserted separately into the same recombinant baculovirus at the egt, p10, and polyhedron gene sites, respectively. The packaging efficiency of rAAV would thus be greater than that when using two or three recombinant baculoviruses (Galibert & Merten, 2011) .