scholarly journals Designing, Construction and Expression of a Recombinant Fusion Protein Comprising the Hepatitis E Virus ORF2 and Rotavirus NSP4 in the Baculovirus Expression System

2016 ◽  
Vol 9 (11) ◽  
Author(s):  
Manoochehr Makvandi ◽  
Ali Teimoori ◽  
Niloofar Neisi ◽  
Alireza Samarbafzadeh
Keyword(s):  
Fusion Protein ◽  
Hepatitis E Virus ◽  
Expression System ◽  
Baculovirus Expression System ◽  
Hepatitis E ◽  
Recombinant Fusion Protein ◽  
Baculovirus Expression

scholarly journals Cloning and expression of hepatitis E virus ORF2 as an immunogen protein in baculovirus expression system

2018 ◽  
Vol 5 (1) ◽  
pp. 23-26
Author(s):  
SA Sadeghi ◽  
M Shahanaghi ◽  
MR Aghasadeghi ◽  
F Motevalli ◽  
MR Amiran ◽  
...  
Keyword(s):  
Hepatitis E Virus ◽  
Expression System ◽  
Baculovirus Expression System ◽  
Hepatitis E ◽  
Cloning And Expression ◽  
Baculovirus Expression

scholarly journals Expression, purification and structural analysis of functional GABA transporter 1 using the baculovirus expression system

2017 ◽  
Vol 13 ◽  
pp. 874-882 ◽  
Author(s):  
Jing Hu ◽  
Chris Weise ◽  
Christoph Böttcher ◽  
Hua Fan ◽  
Jian Yin
Keyword(s):  
Fusion Protein ◽  
Fluorescent Protein ◽  
Expression System ◽  
Baculovirus Expression System ◽  
Size Exclusion ◽  
Coupled Transport ◽  
Gaba Transporter ◽  
Gfp Fusion Protein ◽  
Baculovirus Expression ◽  
Gaba Transporter 1

The γ-aminobutyric acid (GABA) transporter 1 (GAT1) belongs to a family of Na+ and Cl−-coupled transport proteins and possesses 12 putative transmembrane domains. To perform structural analyses of the GAT1 protein, the GAT1/green fluorescent protein (GFP) fusion protein was functionally expressed in insect Sf9 cells by the BAC-TO-BAC® baculovirus expression system. A two-step procedure to purify the GAT1/GFP fusion protein from insect Sf9 cells has been established and involves immunoaffinity chromatography using self-prepared anti-GFP antibodies and size-exclusion fast protein liquid chromatography (SE-FPLC). A yield of 200–300 μg of the GAT1/GFP protein could be purified from 400–600 mL of infected Sf9 cells. The purified protein was analyzed by transmission electron microscopy (TEM), which revealed that the GAT1/GFP fusion protein was isolated in its monomeric form.

scholarly journals Pigs Immunized with the Virus-like Particle Vaccine Are Protected against the Hepatitis E-3 Virus

Vaccines ◽  
2021 ◽  
Vol 9 (11) ◽  
pp. 1265
Author(s):  
Hyeon-Jeong Go ◽  
Byung-Joo Park ◽  
Hee-Seop Ahn ◽  
Dong-Hwi Kim ◽  
Da-Yoon Kim ◽  
...  
Keyword(s):  
Expression System ◽  
Vaccine Dose ◽  
Baculovirus Expression System ◽  
Hepatitis E ◽  
Positive Control ◽  
Negative Control ◽  
Fecal Shedding ◽  
Viral Shedding ◽  
Baculovirus Expression ◽  
Virus Like Particle

In this study, we generated the HEV virus-like particle (VLP) vaccine expressing 239 amino acids (367–605 aa) of the HEV-3 ORF2 using the baculovirus expression system. The HEV-3-239-VLP vaccine efficacy was evaluated by dividing 12 pathogen-free pigs into four groups: negative control, positive control, 100 μg VLP-, and 200 μg VLP-vaccinated groups for 10 weeks. The pigs in either of the vaccinated groups were administered the corresponding first and booster doses on weeks 0 and 2. At week 4, the positive control and two vaccinated groups were challenged with 106 HEV-3 genomic equivalent copies; viremia and fecal shedding of the virus were identified in pigs in the positive control and 100 μg VLP-vaccinated pigs showed transient viremia and fecal viral shedding. However, no viruses were detected in the serum or fecal samples of the 200 μg VLP-vaccinated pigs. The 100 and 200 μg VLP-vaccinated pigs had significantly higher (p < 0.01) anti-HEV antibodies than the negative control pigs from weeks 6–10 with normal levels of liver enzymes. The 200 μg VLP-vaccinated pigs showed statistically less liver tissue fibrosis (p < 0.05) than that of the positive control pigs. Thus, the novel baculovirus expression system-generated VLP vaccine dose-dependently protects against HEV-3 challenge and may be useful in other animal species, including humans.

Production of a urokinase plasminogen activator-IgG fusion protein (uPA-IgG) in the baculovirus expression system

Gene ◽  
1997 ◽  
Vol 190 (1) ◽  
pp. 139-144 ◽  
Author(s):  
Thomas A. Kost ◽  
Diane M. Ignar ◽  
William C. Clay ◽  
John Andrews ◽  
Jeremy D. Leray ◽  
...  
Keyword(s):  
Fusion Protein ◽  
Plasminogen Activator ◽  
Expression System ◽  
Baculovirus Expression System ◽  
Urokinase Plasminogen Activator ◽  
Baculovirus Expression

scholarly journals Characterization of self-assembled virus-like particles of rat hepatitis E virus generated by recombinant baculoviruses

2011 ◽  
Vol 92 (12) ◽  
pp. 2830-2837 ◽  
Author(s):  
Tian-Cheng Li ◽  
Kumiko Yoshimatsu ◽  
Shumpei P. Yasuda ◽  
Jiro Arikawa ◽  
Takaaki Koma ◽  
...  
Keyword(s):  
Hepatitis E Virus ◽  
Expression System ◽  
Recombinant Baculovirus ◽  
Baculovirus Expression System ◽  
Hepatitis E ◽  
Serum Samples ◽  
Electron Microscopic ◽  
Rat Serum ◽  
Wild Rats ◽  
Self Assembled

Hepatitis E virus (HEV) is a causative agent of hepatitis E. Recently, a novel hepatitis E-like virus was isolated from Norway rats in Germany. However, the antigenicity, pathogenicity and epidemiology of this virus are unclear because of the lack of a cell-culture system in which to grow it. In this study, an N-terminally truncated ORF2 protein was expressed in insect Tn5 cells using a recombinant baculovirus expression system and a large amount of 53 kDa protein was expressed and efficiently released into the supernatant. Electron microscopic analyses of the purified 53 kDa protein revealed that the protein self-assembled into two types of empty HEV-like particles (rat HEVLPs). The smaller rat HEVLPs were estimated to be 24 nm in diameter, which is similar to the size of genotype G1, G3 and G4 HEVLPs. The larger rat HEVLPs were estimated to measure 35 nm in diameter, which is similar to the size of native rat HEV particles. An ELISA to detect antibodies was established using rat HEVLPs as the antigens, which demonstrated that rat HEVLPs were cross-reactive with G1, G3 and G4 HEVs. Detection of IgG and IgM antibodies was performed by examination of 139 serum samples from wild rats trapped in Vietnam, and it was found that 20.9 % (29/139) and 3.6 % (5/139) of the samples were positive for IgG and IgM, respectively. In addition, rat HEV RNA was detected in one rat serum sample that was positive for IgM. These results indicated that rat HEV is widespread and is transmitted among wild rats.

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