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

Huijuan Yin; Qin Yao; Zhongjian Guo; Fang Bao; Wei Yu; Jun Li; Keping Chen

doi:10.1016/S1673-8527(08)60033-8

Volume 35 Issue 4

Apr. 2008

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Article Navigation > Journal of Genetics and Genomics > 2008 > 35(4): 239-244

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

doi: 10.1016/S1673-8527(08)60033-8

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

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Corresponding author: E-mail address: yaoqin@ujs.edu.cn (Qin Yao)
Received Date: 2007-09-04
Accepted Date: 2007-10-18
Rev Recd Date: 2007-10-08

Available Online: 2008-04-24

Publish Date: 2008-04-20

Abstract

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

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References(21)

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