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Efficient production of canine interferon-alpha in silkworm Bombyx mori by use of a BmNPV/Bac-to-Bac expression system

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We exploited the silkworm Bombyx mori for the production of recombinant canine interferon-alpha (CaIFN-α). The recombinant baculovirus harboring canine interferon gene was rapidly generated by the BmNPV/Bac-to-Bac system that was recently developed. In B. mori-derived cell lines, the expression of the recombinant protein reached maximal levels around 72–96 h post-infection. For the isolation of the expressed recombinant protein from B. mori larvae, the whole bodies of the infected larvae were homogenized, and the expressed protein was purified by affinity chromatography. Based on the fact that the recombinant CaIFN-α showed two bands on the sodium dodecyl sulfate polyacrylamide gel electrophoresis pattern, the expressed protein was thought to be glycosylated. The rCaIFN-α yield was about 528 μg per larva, showing that the expression in silkworm was successful. Furthermore, the recombinant protein was proven to be able to inhibit the infection of Madin–Darby canine kidney cells by the vesicular stomatitis virus, indicating that it is biologically active in vitro. The method established in this study provides an efficient way to produce a large amount of CaIFN-α and paves the way for further utilization of this protein as a therapeutic agent or vaccine adjuvant in dogs.

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This work was supported by the National High-tech R&D Program (2007AA10Z159), Key International Cooperation Programme of Zhejiang (2006C24015), and a grant of the New Century Training Programme Foundation for the Talents (NCET-06-0524), Ministry of Education, China.

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Correspondence to Wu Xiaofeng.

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Na, Z., Huipeng, Y., Lipan, L. et al. Efficient production of canine interferon-alpha in silkworm Bombyx mori by use of a BmNPV/Bac-to-Bac expression system. Appl Microbiol Biotechnol 78, 221–226 (2008). https://doi.org/10.1007/s00253-007-1296-y

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  • Canine interferon-α
  • BmNPV/Bac-to-Bac expression system
  • Silkworm