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Baculovirus Expression and Potential Diagnostic Application of the Gp51 Envelope Glycoprotein of Genetic Mutants of the Bovine Leukaemia Virus

INTRODUCTION: Field isolates of bovine leukaemia virus (BLV) show the presence of a few amino acid substitutions in major conformational G and H epitopes on surface glycoprotein gp51. Potentially, these substitutions can affect the 3D structure of these epitopes leading to their diminished immunorea...

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Autores principales: Rola-Łuszczak, Marzena, Grabowska, Agnieszka, Szewczyk, Bogusław, Kuźmak, Jacek
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Sciendo 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6458546/
https://www.ncbi.nlm.nih.gov/pubmed/30989129
http://dx.doi.org/10.2478/jvetres-2019-0020
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author Rola-Łuszczak, Marzena
Grabowska, Agnieszka
Szewczyk, Bogusław
Kuźmak, Jacek
author_facet Rola-Łuszczak, Marzena
Grabowska, Agnieszka
Szewczyk, Bogusław
Kuźmak, Jacek
author_sort Rola-Łuszczak, Marzena
collection PubMed
description INTRODUCTION: Field isolates of bovine leukaemia virus (BLV) show the presence of a few amino acid substitutions in major conformational G and H epitopes on surface glycoprotein gp51. Potentially, these substitutions can affect the 3D structure of these epitopes leading to their diminished immunoreactivity. The aim of this study was to express three gp51 glycoproteins carrying mutated epitopes as recombinant baculovirus proteins in insect cells to test their immunoreactivity with bovine sera. MATERIAL AND METHODS: Env gene chimeras encoding mutated epitopes G and H in the env backbone of BLV FLK strain were constructed, cloned into pFastBac1 vector, and expressed in baculovirus. RESULTS: The presence of recombinant gp51 protein in Sf9 insect cells was confirmed using monoclonal antibodies. ELISA tests were developed to check the immunoreactivity of recombinant protein with bovine sera. CONCLUSION: Recombinant gp51 proteins with altered G and H epitopes can be used for further studies to analyse the serological response of bovine sera towards BLV antigenic variants.
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spelling pubmed-64585462019-04-15 Baculovirus Expression and Potential Diagnostic Application of the Gp51 Envelope Glycoprotein of Genetic Mutants of the Bovine Leukaemia Virus Rola-Łuszczak, Marzena Grabowska, Agnieszka Szewczyk, Bogusław Kuźmak, Jacek J Vet Res Research Article INTRODUCTION: Field isolates of bovine leukaemia virus (BLV) show the presence of a few amino acid substitutions in major conformational G and H epitopes on surface glycoprotein gp51. Potentially, these substitutions can affect the 3D structure of these epitopes leading to their diminished immunoreactivity. The aim of this study was to express three gp51 glycoproteins carrying mutated epitopes as recombinant baculovirus proteins in insect cells to test their immunoreactivity with bovine sera. MATERIAL AND METHODS: Env gene chimeras encoding mutated epitopes G and H in the env backbone of BLV FLK strain were constructed, cloned into pFastBac1 vector, and expressed in baculovirus. RESULTS: The presence of recombinant gp51 protein in Sf9 insect cells was confirmed using monoclonal antibodies. ELISA tests were developed to check the immunoreactivity of recombinant protein with bovine sera. CONCLUSION: Recombinant gp51 proteins with altered G and H epitopes can be used for further studies to analyse the serological response of bovine sera towards BLV antigenic variants. Sciendo 2019-03-22 /pmc/articles/PMC6458546/ /pubmed/30989129 http://dx.doi.org/10.2478/jvetres-2019-0020 Text en © 2019 M. Rola-Łuszczak et al., published by Sciendo http://creativecommons.org/licenses/by-nc-nd/3.0 This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.
spellingShingle Research Article
Rola-Łuszczak, Marzena
Grabowska, Agnieszka
Szewczyk, Bogusław
Kuźmak, Jacek
Baculovirus Expression and Potential Diagnostic Application of the Gp51 Envelope Glycoprotein of Genetic Mutants of the Bovine Leukaemia Virus
title Baculovirus Expression and Potential Diagnostic Application of the Gp51 Envelope Glycoprotein of Genetic Mutants of the Bovine Leukaemia Virus
title_full Baculovirus Expression and Potential Diagnostic Application of the Gp51 Envelope Glycoprotein of Genetic Mutants of the Bovine Leukaemia Virus
title_fullStr Baculovirus Expression and Potential Diagnostic Application of the Gp51 Envelope Glycoprotein of Genetic Mutants of the Bovine Leukaemia Virus
title_full_unstemmed Baculovirus Expression and Potential Diagnostic Application of the Gp51 Envelope Glycoprotein of Genetic Mutants of the Bovine Leukaemia Virus
title_short Baculovirus Expression and Potential Diagnostic Application of the Gp51 Envelope Glycoprotein of Genetic Mutants of the Bovine Leukaemia Virus
title_sort baculovirus expression and potential diagnostic application of the gp51 envelope glycoprotein of genetic mutants of the bovine leukaemia virus
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6458546/
https://www.ncbi.nlm.nih.gov/pubmed/30989129
http://dx.doi.org/10.2478/jvetres-2019-0020
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