Expression of Green Fluorescent Protein in Both Spodoptera frugiperda Cells and Bombyx mori Larvae by Ac-Bm Hybrid Virus

  • Jin, Byung-Rae (The Research Center for New Bio-Materials in Agriculture, and Division of Applied Biology and Chemistry, College of Agriculture & Life Sciences, Seoul National University) ;
  • Yoon, Hyung-Joo (National Sericulture & Entomology Research Institute, RDA) ;
  • Yun, Eun-Young (National Sericulture & Entomology Research Institute, RDA) ;
  • Kang, Seok-Woo (National Sericulture & Entomology Research Institute, RDA) ;
  • Cho, Eun-Sook (College of Natural Resources and Life Science, Dong-A university) ;
  • Kang, Seok-Kwon (The Research Center for New Bio-Materials in Agriculture, and Division of Applied Biology and Chemistry, College of Agriculture & Life Sciences, Seoul National University)
  • Published : 1998.09.30

Abstract

We have expressed GFP in Sf9 and Bm5 cells or Bombyx mori larvae by using Ac-Bm hybrid virus capable of replicating in both Bm5 and Sf9 cells. Genomic DNA of Ac-Bm hybrid virus expressing ${\beta}$-galactosidase was cotransfected with baculovirus transfer vector containing GFP gene, pBacPAK-GFP in Sf9 cells. The Ac-Bm hybrid virus harboring GFP was named as Ac-Bm hybrid virus-GFP. The Ac-Bm hybrid virus-GFP-infected insect cells were easily selected by detecting the emission of GFP from each well of cell culture dish on the UV illuminator. GFP produced by Ac-Bm hybrid virus-GFP in Sf9 and Bm5 cells or B. mori larvae was confirmed by SDS-PAGE and Western blot analysis using GFP antibody. In addition, B. mori larvae infected with Ac-Bm hybrid virus-GFP was apparently appeared fluorescence from the whole body at S days postinoculation. The fluorescence of GFP from the hemolymph and fat body of B. mori larvae infected with Ac-Bm hybrid virus-GFP was also observed by fluorescence microscope. In conclusion, our results demonstrated that in baculovirus expression vector system, use of Ac-Bm hybrid virus have an additional advantage of expanded host range for producing recombinant proteins.

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