Protein Expression of the Human Norovirus Capsid Gene using the Baculovirus Expression System

Ji-Young Jin; Chul Jong Park; Seung-Won Park; Soon-Young Paik

doi:10.4167/jbv.2011.41.3.183

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Jin, Park, Park, and Paik: Protein Expression of the Human Norovirus Capsid Gene using the Baculovirus Expression System

Original Article

J Bacteriol Virol 2011;41(3):183-187.

Published online: 18 January 2011

DOI: https://doi.org/10.4167/jbv.2011.41.3.183

Protein Expression of the Human Norovirus Capsid Gene using the Baculovirus Expression System

Ji-Young Jin^1,^†, Chul Jong Park^2,^†, Seung-Won Park³, Soon-Young Paik^1,^∗

¹Department of Microbiology, College of Medicine, The Catholic University, Seoul, Korea

²Department of Dermatology, College of Medicine, The Catholic University, Seoul, Korea

³Department of Agricultural Biology, National Academy of Agricultural Science, Rural Development Association, Suwon, Korea

^∗Corresponding author: Soon-Young Paik. Department of Microbiology, College of Medicine, The Catholic University of Korea, Seoul, 137-701, Korea. Phone: +82-2-2258-7342, Fax: +82-2-535-6473 e-mail: paik@catholic.ac.kr

^† Equal contributors

^∗∗ This work was supported by the Mid-career Researcher Program (2008-0061575) and the Basic Science Research Program (2009-0062720) through an NRF grant funded by the MEST.

Revised 17 May 201126 August 2011 Accepted 29 August 2011

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Human norovirus (HuNoV) is the major etiological agent of nonbacterial gastroenteritis worldwide. However, due to the absence of a rapid and sensitive diagnostic system, detection and monitoring have been limited. The HuNoV genome is composed of three open reading frames (ORFs). And major capsid protein, ORF2, is designated as a viral protein 1 (VP1). In this study, the baculovirus expression system was used for expression of the HuNoV capsid protein, VP1. Recombinant baculoviruses can be used as potent tools in HuNoV studies.

Keywords: Human norovirus, Baculovirus expression system

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Figure 1.

Amplified Fragments of ORF2 S-P1 (543 bp), ORF2 epi (1,224 bp), and ORF2 P2 (347 bp) of the HuNoV capsid gene by RT-PCR (M: DNA size Marker).

Figure 2.

The Gateway cloning strategy. The process comprises of two recombination processes; TOPO cloning reaction (A) and LR recombinant reaction (B).

Figure 3.

Analysis of expression of recombinant protein from insect cells by SDS-PAGE; ORF2 epi (54 kDa), ORF2 S-P1 (29 kDa), and ORF2 P2 (23.4 kDa).

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