Expression of VP2 Gene of Infectious Bursal Disease Virus in Insect Cells
Author(s): YANG Feng, LI Xian-wei, SUN Min-hua, YAN Fu-qiang, WEI Qing-lan, XIANG Bin, REN Tao, Key Laboratory of New Drug Discovery, Ministry of Agriculture, College of Veterinary Medicine, South China Agricultural University
Pages: 65-69
Year: 2013 Issue: 9
Journal: China Animal Husbandry & Veterinary Medicine
Keyword: infectious bursal disease virus; VP2 gene; insect expression vector;
Abstract: The VP2 gene of very virulent infectious bursal disease virus( vvIBDV) JM-1 /10 strain was placed behind the CMV promoter of pcDNA-3. 1( +). The CMV-VP2 gene was cloned into the baculovirus expression system pFastBacTMDual to build pFastCMV-VP2. Transformed it to Escherichia coli DH10Bac. Recombinant baculovirus expression plasmid Bacmid-CMV-VP2 was screened. Bacmid-CMV-VP2 was used to be transfected into Sf9 insect cells to get the recombinant baculovirus vBac-CMV-VP2 and transduced into BHK-21 cells. After 48 to 72 hours,indirect immunofluorescence assay( IFA) was conducted to detect specific fluorescence. Western blotting analysis result showed target protein was expressed. The prepared recombinant vBac-CMV-VP2 could not only be expressed in insect cells,but also expressed in mammalian cells.
Pages: 65-69
Year: 2013 Issue: 9
Journal: China Animal Husbandry & Veterinary Medicine
Keyword: infectious bursal disease virus; VP2 gene; insect expression vector;
Abstract: The VP2 gene of very virulent infectious bursal disease virus( vvIBDV) JM-1 /10 strain was placed behind the CMV promoter of pcDNA-3. 1( +). The CMV-VP2 gene was cloned into the baculovirus expression system pFastBacTMDual to build pFastCMV-VP2. Transformed it to Escherichia coli DH10Bac. Recombinant baculovirus expression plasmid Bacmid-CMV-VP2 was screened. Bacmid-CMV-VP2 was used to be transfected into Sf9 insect cells to get the recombinant baculovirus vBac-CMV-VP2 and transduced into BHK-21 cells. After 48 to 72 hours,indirect immunofluorescence assay( IFA) was conducted to detect specific fluorescence. Western blotting analysis result showed target protein was expressed. The prepared recombinant vBac-CMV-VP2 could not only be expressed in insect cells,but also expressed in mammalian cells.
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