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Wu Carol P.,Chang Chia‐Jung,Li Cheng‐Hsun,Wu Yueh‐Lung 한국곤충학회 2021 Entomological Research Vol.51 No.4
The baculovirus expression system has been widely used for industrial-scale production of recombinant proteins. These recombinant proteins are expressed from recombinant baculoviruses carrying an expression cassette for a particular protein. Even though the baculovirus genome can accommodate large fragments of foreign genetic material, it has been shown that defective interfering particles (DIs) are generated over many passages of the recombinant virus. Defective interfering particles can affect the stability of the inserted gene, which can result in sub-optimal expression of the recombinant protein; this can, therefore, have a significant impact on production. The stability of exogenous genes in recombinant viruses over serial passage has not yet been investigated. As such, the present study set out to assess the effect of serial passage on the stability of the expression of an exogenous gene inserted into a recombinant baculovirus and its subsequent recombinant protein yield. To achieve this, a recombinant baculovirus carrying the classical swine fever virus E2 gene was serially passaged up to 20 times. The results showed that the E2 gene expression by the recombinant baculovirus was stable for up to 10 passages without any effect on recombinant E2 production in insect cells.
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Analyses of the transcriptome of Bombyx mori cells infected with either BmNPV or AcMNPV
Yu-Wei Chen,Carol-P Wu,Tsai-Chin Wu,Yueh-Lung Wu 한국응용곤충학회 2018 Journal of Asia-Pacific Entomology Vol.21 No.1
Baculoviruses are insect-specific DNA viruses with a restricted host range. Two examples of such includeAutographa californica nucleopolyhedrovirus (AcMNPV) and Bombyx mori nucleopolyhedrovirus (BmNPV). AcMNPV is a commercially available and widely used baculovirus prototype which can infect 39 species in 13families. On the other hand, BmNPV is a major pathogen of silkworms which has developed high host specificityto Bombyx mori. Coincidentally, AcMNPV and BmNPV are highly homologous, but share no overlapping hostrange on a genomic level. In fact, these two similar viruses have extremely different infection outcomes inBombyx mori. We hypothesized that the determination of host specificity may depend on virus-host interactionsand several genes may be involved in determining host specificity. Therefore, we used next generation sequencing(NGS) to analyze transcriptome responses of hosts to these viruses. A transcriptome library was constructed,annotated, and grouped after sequence assembly. The comparison of gene expressions showed severalsignificant differences in the gene expression profiles of BmNPV and AcMNPV, especially in cases where genesinvolved with immune responses were verified by RNA interference. In addition, studies have shown that smallRNA plays a role in virus-host interaction, and further determines host specificity. Therefore, we screenedmicroRNA induced by AcMNPV infection, and then combined NGS data from cellular gens to predict possibleregulation networks. The manipulation of virus-host specificity can provide a breakthrough for the application ofbaculovirus in protein expression systems and the development of bio-control agents.
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