Springer Nature is making Coronavirus research free. View research | View latest news | Sign up for updates

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

  • 264 Accesses

  • 18 Citations

Abstract

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.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

References

  1. Belardelli F, Ferrantini M, Proietti E, Kirkwood JM (2002) Interferon-alpha in tumor immunity and immunotherapy. Cytokine Growth Factor Rev 13(2):119–134

  2. Bogdan C (2000) The function of type I interferons in antimicrobial immunity. Curr Opin Immunol 12(4):419–424

  3. Cuiping C, Xingmeng L, Na Z, Huipeng Y, Xiaofeng W, Yeping T (2006) Development of a rapid and efficient BmNPV baculovirus expression system for application in mulberry silkworm, Bombyx mori. Curr Sci 91(12):1692–1697

  4. Decker T, Muller M, Stockinger S (2005) The yin and yang of type I interferon activity in bacterial infection. Nat Rev Immunol 5(9):675–687

  5. Fumiyoshi O, Masahiro S, Takayoshi I, Naoyo O, Katsushige Y (2000) Production of canine IFN-γ in silkworm by recombinant baculovirus and characterization of the product. J Interferon Cytokine Res 20:1015–1022

  6. Giovanna G, Rossella L, Mirella D’A, Vincenzo L, Federica M, Ottavio P, Mirella L, Mara M, Aldo R, Marialuisa Z, Monica M (2006) Expression of bioactive recombinant bovine interferon-γ using baculovirus. New Microbiol 29:19–24

  7. Himmler A, Hauptmann R, Adolf GR, Swetly P (1987) Structure and expression in Escherichia coli of canine interferon-alpha genes. J Interferon Res 7(2):173–183

  8. Jeefrey LC, Charles SC (1996) Protein engineering. Chapter 7: insect cell expression technology. Wiley, New York, pp 183–218

  9. Ohya K, Matsumura T, Itchoda N, Ohashi K, Onuma M, Sugimoto C (2005) Ability of orally administered IFN-alpha-containing transgenic potato extracts to inhibit Listeria monocytogenes infection. J Interferon Cytokine Res 25(8):459–466

  10. Ruttanapumma R, Anzai M, Takegawa M, Okamura M, Maehara N, Sato K, Yoshioka K, Itoh A, Nakamura M, Takehara K (2006) High level expression of biologically active canine interferon-alpha subtype 4 using a baculovirus. J Vet Med Sci 68(12):1347–1349

  11. Taira O, Watanugi I, Hagiwara Y, Takahashi M, Arai S, Sato H, Maehara N (2005) Cloning and expression of canine interferon-alpha genes in Escherichia coli. J Vet Med Sci 67(10):1059–1062

  12. Samuel CE (2001) Antiviral actions of interferons. Clin Microbiol Rev 14(4):778–809

  13. Xiaofeng W, Cao C, Xu Y, Lu X (2004a) Construction of a host range-expanded hybrid baculovirus of BmNPV and AcNPV, and knockout of cysteinase gene for more efficient expression. Sci China C Life Sci 34(2):156–164

  14. Xiaofeng W, Yin Z, Cao C, Ling H, Xingmeng L (2004b) Expression of human VEGF165 in silkworm (Bombyx mori L.) by using a recombinant baculovirus and its bioactivity assay. J Biotechnol 111(3):253–261

Download references

Acknowledgment

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.

Author information

Correspondence to Wu Xiaofeng.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

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

Download citation

Keywords

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