Production of capsid proteins of rat hepatitis E virus in Escherichia coli and characterization of self-assembled virus-like particles

被引：6
|
作者：
Kobayashi, Tominari [1 ]
Takahashi, Masaharu [1 ]
Ohta, Satoshi [2 ]
Nagashima, Shigeo [1 ]
Primadharsini, Putu Prathiwi [1 ]
Mulyanto [3 ,4 ]
Kunita, Satoshi [5 ]
Murata, Kazumoto [1 ]
Okamoto, Hiroaki [1 ]
机构：
[1] Jichi Med Univ, Sch Med, Dept Infect & Immun, Div Virol, 3311-1 Yakushiji, Shimotsuke, Tochigi 3290498, Japan
[2] Jichi Med Univ, Sch Med, Dept Biochem, Div Struct Biochem, Shimotsuke, Tochigi 3290498, Japan
[3] West Nusa Tenggara Hepatitis Lab, Mataram, Indonesia
[4] Univ Mataram, Fac Med, Immunobiol Lab, Mataram, Indonesia
[5] Jichi Med Univ, Sch Med, Ctr Expt Med, Shimotsuke, Tochigi 3290498, Japan
来源：
VIRUS RESEARCH | 2021年 / 302卷
关键词：
Rat hepatitis E virus; Capsid protein; Virus-like particle; Escherichia coli; Assembly in vitro; MONOCLONAL-ANTIBODIES; WILD RATS; ORF3; PROTEIN; PIG-LIVER; E VACCINE; RECOMBINANT; HEV; TRANSMISSION; IDENTIFICATION; GENOTYPE;
D O I：
10.1016/j.virusres.2021.198483
中图分类号：
Q93 [微生物学];
学科分类号：
071005 ; 100705 ;
摘要：
Rat hepatitis E virus (HEV) has been isolated from wild rats worldwide and the potential of zoonotic transmission has been documented. Escherichia coli (E. coli) is utilized as an effective system for producing HEV-like particles. However, the production of rat HEV ORF2 proteins in E. coli forming virus-like particles (VLPs) has not yet been reported. In this study, nine rat HEV ORF2 proteins of the ratELOMB-131L strain with truncated N- and C-termini (amino acids 339-594, 349-594, 351-594, 354-594, 357-594, 357-599, 357-604, 357-609, and 357-614 of ORF2 protein) were expressed in E. coli and the 357-614 protein self-assembled most efficiently. A bioanalyzer showed that the purified 357-614 protein has a molecular weight of 33.5 kDa and a purity of 93.2%. Electron microscopy revealed that the purified 33.5 kDa protein formed VLPs with a diameter of 21-52 (average 32) nm, and immunoelectron microscopy using an anti-rat HEV ORF2 monoclonal antibody (TA7014) indicated that the observed VLPs were derived from rat HEV ORF2. The VLPs attached to and entered the PLC/PRF/5 cells and blocked the neutralization of rat HEV by TA7014, suggesting that the VLPs possess the antigenic structure of infectious rat HEV particles. In addition, rat HEV VLPs showed high immunogenicity in mice. The present results would be useful for future studies on the development of VLP-based vaccines for HEV prevention in a rat model and for the prevention of rat HEV infection in humans.
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页数：10
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