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Cloning and Expression of Spodoptera litura nucleopolyhedrovirus
gp41 Gene in Escherichia coli and Preparation of Its Antibody |
| PAN Li-Jing LI Zhao-Fei YIN Chong LV Lei PANG Yi ** |
| （State Key Laboratory for Biocontrol, Zhongshan University, Guangzhou 510275, China） |
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Abstract Abstract: GP41, a major glycoprotein, has been identified in the Occlusion-derived virus（ODV）of baculoviruses, which was required for the egress of nucleocapsids from the nucleus in the pathway of the Budded virus（BV）synthesis. The ORF of Spodoptera litura nucleopolyhedro virus（SpltMNPV）gp41 gene was obtained by PCR method from SpltMNPV genomic DNA. The PCR product was cloned into pMD18-T vector to get the recombinant plasmid（pT-gp41）. The gp41 gene was recombined in vitro with prokaryotic expression vector pQE30 and transformed into Escherichia.coli M15 [pREP4]. The M15 [pREP4] strain, containing gp41 recombinant plasmid, expressed a 37.9 kD 6×His-tag fusion protein after induction with 1 mmol/L IPTG (isopropylthio-β-D-galactoside). The fusion protein was purified with Ni-NTA resin column and used as the immunogen to raise a GP41-specific antibody. Western blotting analysis indicated that the antibody was suitable to be used for further analysis of GP41 protein.
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Received: 01 January 1900
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Corresponding Authors:
PANG Y
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