Skip to main content Accessibility help
×
Home

Cloning and expression of Spodoptera litura nucleopolyhedrovirus gp41 gene in Escherichia coli and preparation of antibody

  • Pan Li-Jing (a1), Li Zhao-Fei (a1), Yin Chong (a1), Lv Lei (a1) and Pang Yi (a1)...

Abstract

GP41, a major glycoprotein, identified in the occlusion-derived virions (ODV) of baculoviruses, is required for the egress of nucleocapsids from the nucleus in the pathway of budded virion (BV) synthesis. Using the polymerase chain reaction (PCR), the open reading frame (ORF) of Spodoptera litura nucleopolyhedrovirus (SpltMNPV) gp41 gene was obtained 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 kDa 6×His-tag fusion protein after induction with 1 mmol/l isopropylthio-β-d-galactoside (IPTG). The fusion protein was purified with a nickel-nitrilotriacetic acid (Ni–NTA) resin column and used as the immunogen to raise GP41-specific antibody. Western blotting analysis indicated that the antibody was suitable to be used for further analysis of GP41 protein.

Copyright

Corresponding author

*Corresponding author. Email: LS12@zsu.edu.cn

References

Hide All
Herniou, EA, Olszewski, JA, Cory, JS et al. , (2003) The genome sequence and evolution of baculoviruses. Annual Review of Entomology 48: 211234.
Li, GH, Chen, QJ and Pang, Y (1988) Studies on artificial diets for the beet armyworm, Spodoptera exigua. Acta Scientiarum Naturalium Universitatis SunYatSeNi 37 (4): 15.
Miller, LK (1997) Introduction to the baculovirus. In: Miller, LK (editor) The Baculovirus. New York: Plenum Press, pp. 16.
Olszewski, J and Miller, LK (1997) A role for baculovirus GP41 in budded virus production. Virology 33 (2): 292301.
O'Reilly, DR, Miller, LK and Luckow, VA (1992) Baculovirus Expression Vectors – A Laboratory Manual. New York: W.H. Freeman and Co., pp. 139179.
Pang, Y, Yu, JX, Wang, LH et al. , (2001) Sequence analysis of the Spodoptera litura multicapsid nucleopolyhedrovirus genome. Virology 287: 391404.
Sambrook, J, Fritsch, EF and Maniatis, T (1989) Molecular Cloning: A Laboratory Manual. New York: Cold Spring Harbor Laboratory Press.
Volkman, LE (1986) The 64K envelope protein of budded Autographa californica nuclear polyhedrosis virus. Current Topics in Microbiology and Immunology 131: 103118.
Whitford, M and Faulkner, P (1992) A structural polypeptide of the baculovirus Autographa californica nuclear polyhedrosis virus contains O-linked N-acetylglucosamine. Journal of Virology 66: 33243329.
Xie, WD, Qu, SR, Pang, Y et al. , (1988) Establishment of a cell line from Spodoptera litura and its susceptibility to nuclear polyhedrosis viruses. Acta Scientiarum Naturalium Universitatis SunYatSeNi 4: 113116.

Keywords

Cloning and expression of Spodoptera litura nucleopolyhedrovirus gp41 gene in Escherichia coli and preparation of antibody

  • Pan Li-Jing (a1), Li Zhao-Fei (a1), Yin Chong (a1), Lv Lei (a1) and Pang Yi (a1)...

Metrics

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed