TY - JOUR
T1 - Cloning and expression of human rotavirus spike protein, VP8*, in Escherichia coli
AU - Kovacs-Nolan, Jennifer
AU - Sasaki, Erika
AU - Yoo, Dongwan
AU - Mine, Yoshinori
N1 - Funding Information:
We thank Dr. Shigeru Hayakawa (Department of Biochemistry and Food Science, Kagawa University, Japan) for carrying out the N-terminal sequence analysis, the Arkell Research Sation, Poultry Unit (Guelph, Ontario, Canada) for providing the chickens, and the Central Animal Facility (University of Guelph). This work was supported by grants from the Natural Sciences and Engineering Research Council of Canada, Ontario Egg Producers’ Marketing Board, Agriculture and Agri-Food Canada, and the Ontario Ministry of Agriculture, Food and Rural Affairs.
Copyright:
Copyright 2017 Elsevier B.V., All rights reserved.
PY - 2001
Y1 - 2001
N2 - A system for the expression and purification of soluble VP8*, part of the human rotavirus (HRV) spike protein, was established by expressing VP8* as a fusion protein with glutathione S-transferase (GST). VP8 cDNA, from the Wa strain of HRV, was prepared by RT-PCR, cloned into a pUC18 plasmid, and inserted into a pGEX-4T-2 GST fusion vector. The GST-VP8* fusion protein was expressed in Escherichia coli, and the VP8* was purified by Glutathione Sepharose 4B affinity chromatography, yielding 1.8 mg VP8*/L culture. The purified VP8* was used to vaccinate chickens, eliciting antibodies which displayed high neutralization activity against the Wa strain of HRV, suggesting its use for the induction of specific neutralizing antibodies for potential immunotherapeutic applications for the prevention of HRV infection.
AB - A system for the expression and purification of soluble VP8*, part of the human rotavirus (HRV) spike protein, was established by expressing VP8* as a fusion protein with glutathione S-transferase (GST). VP8 cDNA, from the Wa strain of HRV, was prepared by RT-PCR, cloned into a pUC18 plasmid, and inserted into a pGEX-4T-2 GST fusion vector. The GST-VP8* fusion protein was expressed in Escherichia coli, and the VP8* was purified by Glutathione Sepharose 4B affinity chromatography, yielding 1.8 mg VP8*/L culture. The purified VP8* was used to vaccinate chickens, eliciting antibodies which displayed high neutralization activity against the Wa strain of HRV, suggesting its use for the induction of specific neutralizing antibodies for potential immunotherapeutic applications for the prevention of HRV infection.
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U2 - 10.1006/bbrc.2001.4717
DO - 10.1006/bbrc.2001.4717
M3 - Article
C2 - 11302740
AN - SCOPUS:0034812331
SN - 0006-291X
VL - 282
SP - 1183
EP - 1188
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 5
ER -