Expression of Green Fluorescent Protein in Both Spodoptera frugiperda Cells and Bombyx mori Larvae by Ac-Bm Hybrid Virus.
- Author:
Byung Rae JIN
;
Hyung Joo YOON
;
Eun Young YUN
;
Seok Woo KANG
;
Eun Sook CHO
;
Seok Kwon KANG
- Publication Type:Original Article
- Keywords:
Baculovirus;
Expression vector;
Silkworm;
Insect cells
- MeSH:
Animals;
Baculoviridae;
Blotting, Western;
Bombyx*;
Cell Culture Techniques;
DNA;
Electrophoresis, Polyacrylamide Gel;
Fat Body;
Fluorescence;
Hemolymph;
Host Specificity;
Insects;
Larva*;
Recombinant Proteins;
Sf9 Cells;
Spodoptera*
- From:Journal of the Korean Society of Virology
1998;28(3):225-232
- CountryRepublic of Korea
- Language:English
-
Abstract:
We have expressed GFP in Sf9 and Bm5 cells or Bombyx by larvae by using Ac-Bm hybrid virus capable of replicating in both Bm5 and Sf9 cells. Genomic DNA of Ac-Bm hybrid virus expressing P-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 5 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.
