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KMID : 0357419980280030225
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Korean journal of Virology
1998 Volume.28 No. 3 p.225 ~ p.232
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Expression of Green Fluorescent Protein in Both Spodoptera frugiperda Cells and Bombyx mori Larvae by Ac-Bm Hybrid Virus
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Byung Rae Jin
Hyung Joo Yoon/Eun Young Yun/Seok Woo Kang/Eun Sook Cho/Seok Kwon Kang
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Abstract
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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
β-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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KEYWORD
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Baculovirus, Expression vector, Silkworm, Insect cells,
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