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CLC number: Q812

On-line Access: 2012-01-18

Received: 2011-04-20

Revision Accepted: 2011-07-27

Crosschecked: 2012-01-03

Cited: 3

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Journal of Zhejiang University SCIENCE B 2012 Vol.13 No.2 P.111-117


Immobilization of foreign protein into polyhedra of Bombyx mori nucleopolyhedrovirus (BmNPV)

Author(s):  Xing-wei Xiang, Rui Yang, Lin Chen, Xiao-long Hu, Shao-fang Yu, Cui-ping Cao, Xiao-feng Wu

Affiliation(s):  College of Animal Sciences, Zhejiang University, Hangzhou 310058, China

Corresponding email(s):   wuxiaofeng@zju.edu.cn

Key Words:  Bombyx mori nucleopolyhedrovirus, Polyhedra, Foreign protein, Protein immobilization

Xing-wei Xiang, Rui Yang, Lin Chen, Xiao-long Hu, Shao-fang Yu, Cui-ping Cao, Xiao-feng Wu. Immobilization of foreign protein into polyhedra of Bombyx mori nucleopolyhedrovirus (BmNPV)[J]. Journal of Zhejiang University Science B, 2012, 13(2): 111-117.

@article{title="Immobilization of foreign protein into polyhedra of Bombyx mori nucleopolyhedrovirus (BmNPV)",
author="Xing-wei Xiang, Rui Yang, Lin Chen, Xiao-long Hu, Shao-fang Yu, Cui-ping Cao, Xiao-feng Wu",
journal="Journal of Zhejiang University Science B",
publisher="Zhejiang University Press & Springer",

%0 Journal Article
%T Immobilization of foreign protein into polyhedra of Bombyx mori nucleopolyhedrovirus (BmNPV)
%A Xing-wei Xiang
%A Rui Yang
%A Lin Chen
%A Xiao-long Hu
%A Shao-fang Yu
%A Cui-ping Cao
%A Xiao-feng Wu
%J Journal of Zhejiang University SCIENCE B
%V 13
%N 2
%P 111-117
%@ 1673-1581
%D 2012
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1100131

T1 - Immobilization of foreign protein into polyhedra of Bombyx mori nucleopolyhedrovirus (BmNPV)
A1 - Xing-wei Xiang
A1 - Rui Yang
A1 - Lin Chen
A1 - Xiao-long Hu
A1 - Shao-fang Yu
A1 - Cui-ping Cao
A1 - Xiao-feng Wu
J0 - Journal of Zhejiang University Science B
VL - 13
IS - 2
SP - 111
EP - 117
%@ 1673-1581
Y1 - 2012
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1100131

In the late phase of Bombyx mori nucleopolyhedrovirus (BmNPV) infection, a large amount of polyhedra appear in the infected cell nucleolus, these polyhedra being dense protein crystals protecting the incorporated virions from the harsh environment. To investigate whether the foreign protein could be immobilized into the polyhedra of BmNPV, two recombinant baculoviruses were generated by a novel BmNPV polyhedrin-plus (polh+) Bac-to-Bac system, designated as vBmBac(polh+)-enhanced green fluorescent protein (EGFP) and vBmBac(polh+)-LacZ, which can express the polyhedrin and foreign protein simultaneously. Light microscopy analysis showed that all viruses produced polyhedra of normal appearance. Green fluorescence can be apparently detected on the surface of the vBmBac(polh+)-EGFP polyhedra, but not the BmNPV polyhedra. Fluorescence analysis and anti-desiccation testing confirmed that EGFP was embedded in the polyhedra. As expected, the vBmBac(polh+)-LacZ polyhedra contained an amount of LacZ and had a higher β-galactosidase activity. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting were also performed to verify if the foreign proteins were immobilized into polyhedra. This study provides a new inspiration for efficient preservation of useful proteins and development of new pesticides with toxic proteins.

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article


[1]Blissard, G.W., Rohrmann, G.F., 1990. Baculovirus diversity and molecular biology. Annu. Rev. Entomol., 35(1):127-155.

[2]Cao, C.P., Wu, X.F., Zhao, N., Yao, H.P., Lu, X.M., Tan, Y.P., 2006. Development of a rapid and efficient BmNPV baculovirus expression system for application in mulberry silkworm, Bombyx mori. Curr. Sci., 91(12):1692-1697.

[3]Coulibaly, F., Chiu, E., Ikeda, K., Gutmann, S., Haebel, P.W., Schulze-Briese, C., Mori, H., Metcalf, P., 2007. The molecular organization of cypovirus polyhedra. Nature, 446(7131):97-101.

[4]Coulibaly, F., Chiu, E., Gutmann, S., Rajendran, C., Haebel, P.W., Ikeda, K., Mori, H., Ward, V.K., Schulze-Briese, C., Metcalf, P., 2009. The atomic structure of baculovirus polyhedra reveals the independent emergence of infectious crystals in DNA and RNA viruses. PNAS, 106(52):22205-22210.

[5]Hitchman, R.B., Possee, R.D., Crombie, A.T., Chambers, A., Ho, K., Siaterli, E., Lissina, O., Sternard, H., Novy, R., Loomis, K., et al., 2010. Genetic modification of a baculovirus vector for increased expression in insect cells. Cell Biol. Toxicol., 26(1):57-68.

[6]Ijiri, H., Coulibaly, F., Nishimura, G., Nakai, D., Chiu, E., Takenaka, C., Ikeda, K., Nakazawa, H., Hamada, N., Kotani, E., et al., 2009. Structure-based targeting of bioactive proteins into cypovirus polyhedra and application to immobilized cytokines for mammalian cell culture. Biomaterials, 30(26):4297-4308.

[7]Ikeda, K., Nakazawa, H., Shimo-Oka, A., Ishio, K., Miyata, S., Hosokawa, Y., Matsumura, S., Masuhara, H., Belloncik, S., Alain, R., et al., 2006. Immobilization of diverse foreign proteins in viral polyhedra and potential application for protein microarrays. Proteomics, 6(1):54-66.

[8]Ji, X.Y., Sutton, G., Evans, G., Axford, D., Owen, R., Stuart, D.I., 2010. How baculovirus polyhedra fit square pegs into round holes to robustly package viruses? EMBO J., 29(2):505-514.

[9]Keddie, B.A., Aponte, G.W., Volkman, L.E., 1989. The pathway of infection of Autographa californica nuclear polyhedrosis virus in an insect host. Science, 243(4899):1728-1730.

[10]Mori, H., Ito, R., Nakazawa, H., Sumida, M., Matsubara, F., Minobe, Y., 1993. Expression of Bombyx mori cytoplasmic polyhedrosis virus polyhedrin in insect cells by using a baculovirus expression vector, and its assembly into polyhedra. J. Gen. Virol., 74(1):99-102.

[11]Nordström, T., Senkas, A., Eriksson, S., Pontynen, N., Nordstrom, E., Lindqvist, C., 1999. Generation of a new protein purification matrix by loading ceramic hydroxyapatite with metal ions—demonstration with poly-histidine tagged green fluorescent protein. J. Biotechnol., 69(2-3):125-133.

[12]Reid, B.G., Flynn, G.C., 1997. Chromophore formation in green fluorescent protein. Biochemistry, 36(22):6786-6791.

[13]Rohrmann, G.F., 1986. Polyhedrin structure. J. Gen. Virol., 67(8):1499-1513.

[14]Slack, J., Arif, B.M., 2006. The baculoviruses occlusion-derived virus: virion structure and function. Adv. Virus Res., 69:99-165.

[15]Wang, L., Salem, T.Z., Campbell, D.J., Turney, C.M., Kumar, C.M., Cheng, X.W., 2009. Characterization of a virion occlusion-defective Autographa californica multiple nucleopolyhedrovirus mutant lacking the p26, p10 and p74 genes. J. Gen. Virol., 90(7):1641-1648.

[16]Xiang, X., Yang, R., Yu, S., Cao, C., Guo, A., Chen, L., Wu, X., Cui, W., Cenis, J.L., 2010. Construction of a BmNPV polyhedrin-plus Bac-to-Bac baculovirus expression system for application in silkworm, Bombyx mori. Appl. Microbiol. Biotechnol., 87(1):289-295.

[17]Zhang, G., Gurtu, V., Kain, S.R., 1996. An enhanced green fluorescent protein allows sensitive detection of gene transfer in mammalian cells. Biochem. Biophys. Res. Commun., 227(3):707-711.

[18]Zhao, N., Yao, H.P., Lan, L.P., Cao, C.P., Umashankar, M.L., Lu, X.M., Wu, X.F., Wang, B., Cui, W.Z., Cenis, J.L., 2008. Efficient production of canine interferon-α in silkworm Bombyx mori by use of a BmNPV/Bac-to-Bac expression system. Appl. Microbiol. Biotechnol., 78(2):221-226.

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