Expression of non-structural protein NS3 gene of Bombyx mori densovirus (China isolate)

Institute of Life Sciences, Jiangsu University, 301# Xuefu Road, Zhenjiang 212013, China

More Information

Corresponding author: E-mail address: yaoqin@ujs.edu.cn (Qin Yao)

摘要

- /
- /
Abstract: The invertebrate parvovirus Bombyx mori Densonucleosis Virus type 3 (China isolate), named BmDNV-3, is a kind of bidensovirus. It is a new type of virus with unique replication mechanisms. To investigate the effects of the NS3 gene during viral DNA replication, a pair of primers was designed for amplifying NS3 gene of Bombyx mori densovirus (China isolate). Gene NS3 amplified was cloned into a prokaryotic expression vector pET-30a and the donor plasmid pFastBacHTe, respectively. The NS3 protein was expressed inEscherichia coli BL21. The pFastBacHTe-NS3 was transformed to E. coli DH10Bac. The recombinant bacmid baculoviruses (rBacmid-EGFP-NS3) isolated from the white colonies were transfected into BmN-4 cells using a transfection reagent. BmN-4 cells were infected with recombinant virus to express fusion proteins. The expression of fusion protein around 30 kDa in E. coli BL21 was identified by SDS-PAGE, Western blotting, and mass spectrometry. The expressed NS3 protein by B. mori nucleopolyhedrovirus bacmid system was confirmed by Western blotting using an anti-NS3 polyclonal antibody. And about 45 kDa protein was found. The expressed fusion protein was smaller than the expected size of EGFP-NS3, 55 kDa. Western blotting analysis indicated that EGFP-NS3 protein was expressed in infected larvae with smaller molecular size.
- BmDNV /
- NS3 /
- protein expression

HTML全文

参考文献(21)

[1]	Abd-Alla, A., Jousset, F.X., Li, Y. et al. J. Virol., 78 (2004),pp. 790-797
[2]	Bando, H., Choi, H., Ito, Y. et al. Structural analysis on the single-stranded genomic DNAs of the virus newly isolated from silkworm: the DNA molecules share a common terminal sequence Arch. Virol., 124 (1992),pp. 187-193
[3]	Bando, H., Hayakawa, T., Asano, S. et al. Analysis of the genetic information of a DNA segment of a new virus from silkworm Arch. Virol., 140 (1995),pp. 1147-1155
[4]	Bando, H., Kusuda, J., Gojobori, T. et al. Organization and nucleotide sequence of a densovirus genome imply a host-dependent evolution of the parvoviruses J. Virol., 61 (1987),pp. 553-560
[5]	Bodendorf, U., Cziepluch, C., Jauniaux, J.C. et al. Nuclear export factor CRM1 interacts with nonstructural proteins NS2 from parvovirus minute virus of mice J. Virol., 73 (1999),pp. 7769-7779
[6]	Christensen, J., Tattersall, P. Parvovirus initiator protein NS1 and RPA coordinate replication fork progression in a reconstituted DNA replication system J. Virol., 76 (2002),pp. 6518-6531
[7]	Fediere, G., Li, Y., Zadori, Z. et al. Virology, 292 (2002),pp. 299-308
[8]	Hu, Y.C., Luo, Y.L., Ji, W.T. et al. Dual expression of the HA protein of H5N2 avian influenza virus in a baculovirus system J. Virol. Methods., 135 (2006),pp. 43-48
[9]	Lü, H.
[10]	Li, Y., Zadori, Z., Bando, H. et al. J. Gen. Virol., 82 (2001),pp. 2821-2825
[11]	Miller, C.L., Pintel, D.J. Interaction between parvovirus NS2 protein and nuclear export factor Crm1 is important for viral egress from the nucleus of murine cells J. Virol., 76 (2002),pp. 3257-3266
[12]	Nuesch, J.P., Lachmann, S., Corbau, R. et al. J. Virol., 77 (2003),pp. 433-442
[13]	Qian, G., Youmin, C., Qun, Z. Construction of BmDNV-Z gene libarary Science of Sericulture, 13 (1987),pp. 227-229
[14]	Raab, U., Beckenlehner, K., Lowin, T. et al. NS1 protein of parvovirus B19 interacts directly with DNA sequences of the p6 promoter and with the cellular transcription factors Sp1/Sp3 Virology, 293 (2002),pp. 86-93
[15]	Sambrook, J., David, W., Russell, D.W.
[16]	Tijssen, P., Bergoin, M. Densonucleosis viruses constitute an increasingsy diversified subfamily among the parvoviruses Semin. Virol., 6 (1995),pp. 347-355
[17]	Wang, J., Zhang, J., Jiang, H. et al. Nucleotide sequence and genomic organization of a newly isolated densovirus infecting Dendrolimus punctatus J. Gen. Virol., 86 (2005),pp. 2169-2173
[18]	Wang, Y.J., Yao, Q., Chen, K.P. et al. Virus Genes, 35 (2007),pp. 103-108
[19]	Yang, B., Zhang, J., Cai, D. et al. Biochem. Biophys. Res. Commun., 342 (2006),pp. 1188-1196
[20]	Young, P.J., Jensen, K.T., Burger, L.R. et al. Minute virus of mice NS1 interacts with the SMN protein, and they colocalize in novel nuclear bodies induced by parvovirus infection J. Virol., 76 (2002),pp. 3892-3904
[21]	Young, P.J., Jensen, K.T., Burger, L.R. et al. Minute virus of mice small nonstructural protein NS2 interacts and colocalizes with the Smn protein J. Virol., 76 (2002),pp. 6364-6369