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Expression of vip2A(c) gene from Bacillus thuringiensis in insect cells

Published online by Cambridge University Press:  20 March 2007

Shi Yong-Xia ,
Lv Lei ,
Xu Wei ,
Yuan Mei-Jin  and
Pang Yi
Shi Yong-Xia
Affiliation:
State Key Laboratory for Biocontrol, Zhongshan (Sun Yat-sen) University, Guangzhou 510275, China Guangdong Inspection and Quarantine Technology Center, Guangzhou 510623, China
Lv Lei
Affiliation:
State Key Laboratory for Biocontrol, Zhongshan (Sun Yat-sen) University, Guangzhou 510275, China
Xu Wei
Affiliation:
State Key Laboratory for Biocontrol, Zhongshan (Sun Yat-sen) University, Guangzhou 510275, China
Yuan Mei-Jin
Affiliation:
State Key Laboratory for Biocontrol, Zhongshan (Sun Yat-sen) University, Guangzhou 510275, China
Pang Yi*
Affiliation:
State Key Laboratory for Biocontrol, Zhongshan (Sun Yat-sen) University, Guangzhou 510275, China
*
*Corresponding author. E-mail: [email protected]
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Abstract

Recombinant Bac-GV2 DNA was obtained by inserting a fused gfp gene with the Bacillus thuringiensis vip2A(c) gene encoding a possible enzymatic component under the control of the polyhedrin gene promoter of the baculovirus Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV). The Trichoplusia ni cell line TnHi5 was transfected with Bac-GFP and Bac-GV2 DNAs respectively. Fluorescent cells expressing the fusion protein GV2 were much fewer than those expressing green fluorescent protein (GFP) alone, and did not obviously increase in number from 2 to 5 days after transfection. This result showed that the Vip2A fusion protein might have an ADP-ribosylating activity on cell skeleton actin, exerting an influence on the production and diffusion of the budded virus from insect cells.

Keywords

Bacillus thuringiensisvip2A(c) geneADP-ribosylating activityAutographa californica multicapsid nucleopolyhedrovirus
Type
Research Article
Information
Chinese Journal of Agricultural Biotechnology , Volume 3 , Issue 3 , December 2006 , pp. 223 - 229
Copyright
China Agricultural University and Cambridge University Press 2006

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References

Adang, MJ (1991) Bacillus thuringiensis insecticidal crystal protein:gene structure, action and utilization. In: Maramorosch, K (editor) Biotechnology for Biological Control of Pest and Vectors. London: CRC Press, pp. 3–24.Google Scholar
Collier, RJ (1990) Diphtheria toxin: structure and function of a cytocidal protein. In: Moss, J and Vaughan, M (editors) ADP-ribosylating Toxins and G Proteins. Washington DC: American Society for Microbiology, pp. 3–19.Google Scholar
Estruch, JJ, Warren, GW, Mullins, MA, Nye, GJ, Craig, JA and Koziel, MG (1996) Vip3A, a novel Bacillus thuringiensis vegetative insecticidal protein with a wide spectrum of activities against lepidopteran insects. Proceedings of the National Academy of Sciences of the United States of America 93: 5389–5394.CrossRefGoogle ScholarPubMed
Estruch, JJ, Yu, CG, Warren, GW, Desai, NM, Koziel, MG and Nye, GJ (2000) Class of proteins for the control of plant pests. World Intellectual Property Organization Patent WO 6107279.Google Scholar
Gierschik, P (1992) ADP-ribosylation of signal-transducing guanine nucleotide-binding proteins by pertussis toxin. Current Topics in Microbiology and Immunology 175: 69–98.Google ScholarPubMed
Han, S, Craig, JA, Putnam, CD, Carozzi, NB and Trainer, JA (1999) Evolution and mechanism from structures of an ADP-ribosylating toxin and NAD complex. Nature Structural Biology 6: 932–936.Google ScholarPubMed
Han, S, Arvai, AS, Clancy, SB and Trainer, JA (2001) Crystal structure and novel recognition motif of Rho ADP-ribosylating C3 exoenzyme from Clostridium botulinum: structure insights for recognition specificity and catalysis. Journal of Molecular Biology 305: 95–107.CrossRefGoogle ScholarPubMed
Jarvis, DL (1997) Baculovirus expression vector. In: Miller, LK (editor) The Baculoviruses. New York: Plenum, pp. 389–431.CrossRefGoogle Scholar
Luckow, VA, Lee, SC, Barry, GF and Olins, PO (1993) Efficient generation of infectious recombinant baculoviruses by site-specific transposon-mediated insertion of foreign genes into a baculovirus genome propagated in Escherichia coli. Journal of Virology 67: 4566–4579.CrossRefGoogle ScholarPubMed
Sambrook, J, Fritsch, EF and Maniatis, T (1989) Molecular Cloning: A Laboratory Manual. 2nd ed. New York: Cold Spring Harbor Laboratory Press.Google Scholar
Schirmer, J, Just, I and Aktories, K (2002) The ADP-ribosylating mosquitocidal toxin from Bacillus sphaericus: proteolytic activation, enzyme activity, and cytotoxic effects. Journal of Biological Chemistry 277: 11941–11948.CrossRefGoogle ScholarPubMed
Schnepf, E, Crickmore, N, Van Rie, J, et al. (1998) Bacillus thuringiensis and its pesticidal proteins. Microbiology and Molecular Biology Reviews 62: 775–806.CrossRefGoogle Scholar
Shi, YX, Xu, W, Yuan, MJ, Tang, MJ, Chen, JW and Pang, Y (2004) Expression of vip1/vip2 genes in Escherichia coli and Bacillus thuringiensis and the analysis of their signal peptides. Journal of Applied Microbiology 97: 757–765.CrossRefGoogle ScholarPubMed
Stewart, LM, Hirst, M, Lopez Ferber, M, Merryweather, AT, Cayley, PJ and Possee, RD (1991) Construction of an improved baculovirus insecticide containing an insect-spectific toxin gene. Nature 352: 85–88.CrossRefGoogle ScholarPubMed
Tsuge, H, Nagahama, M, Nishimura, H, et al. (2003) Crystal structure and site-directed mutagenesis of enzymatic components from Clostridium perfringens iota-toxin. Journal of Molecular Biology 325: 471–483.CrossRefGoogle ScholarPubMed
Warren, GW, Koziel, MG and Mullins, MA (1998) Auxiliary proteins for enhancing the insecticidal activity of pesticidal proteins. United States Patent 5770696.Google Scholar
Wilde, C, Vogelsgesang, M and Aktories, K (2003) Rho-specific Bacillus cereus ADP-ribosyltransferase C3cer cloning and characterization. Biochemistry 42: 9694–9702.CrossRefGoogle ScholarPubMed