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DNA immunization with a herpes simplex virus 2 bacterial artificial chromosome()

Construction of a herpes simplex virus 2 (HSV-2) bacterial artificial chromosome (BAC) is described. BAC vector sequences were inserted into the thymidine kinase gene of HSV-2 by homologous recombination. DNA from cells infected with the resulting recombinant virus was transformed into E. coli, and...

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Autores principales: Meseda, Clement A., Schmeisser, Falko, Pedersen, Robin, Woerner, Amy, Weir, Jerry P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier Inc. 2004
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7173264/
https://www.ncbi.nlm.nih.gov/pubmed/14972567
http://dx.doi.org/10.1016/j.virol.2003.09.033
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author Meseda, Clement A.
Schmeisser, Falko
Pedersen, Robin
Woerner, Amy
Weir, Jerry P.
author_facet Meseda, Clement A.
Schmeisser, Falko
Pedersen, Robin
Woerner, Amy
Weir, Jerry P.
author_sort Meseda, Clement A.
collection PubMed
description Construction of a herpes simplex virus 2 (HSV-2) bacterial artificial chromosome (BAC) is described. BAC vector sequences were inserted into the thymidine kinase gene of HSV-2 by homologous recombination. DNA from cells infected with the resulting recombinant virus was transformed into E. coli, and colonies containing the HSV-2 BAC (HSV2-BAC) were isolated and analyzed for the expected genotype. HSV2-BAC DNA was infectious when transfected back into mammalian cells and the resulting virus was thymidine kinase negative. When used to immunize mice, the HSV2-BAC DNA elicited a strong HSV-2 specific antibody response that was equal to or greater than live virus immunization. Further, HSV2-BAC immunization was protective when animals were challenged with a lethal dose of virus. The utility of the HSV2-BAC for construction of recombinant virus genomes was demonstrated by elimination of the HSV-2 glycoprotein D (gD) gene. A recombinant HSV-2 BAC with the gD gene deleted was isolated and shown to be incapable of producing infectious virus following transfection unless an HSV gD gene was expressed in a complementing cell line. Immunization of mice with the HSV2 gD-BAC also elicited an HSV-2 specific antibody response and was protective. The results demonstrate the feasibility of DNA immunization with HSV-2 bacterial artificial chromosomes for replicating and nonreplicating candidate HSV-2 vaccines, as well as the utility of BAC technology for construction and maintenance of novel HSV-2 vaccines. The results further suggest that such technology will be a powerful tool for dissecting the immune response to HSV-2.
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spelling pubmed-71732642020-04-22 DNA immunization with a herpes simplex virus 2 bacterial artificial chromosome() Meseda, Clement A. Schmeisser, Falko Pedersen, Robin Woerner, Amy Weir, Jerry P. Virology Article Construction of a herpes simplex virus 2 (HSV-2) bacterial artificial chromosome (BAC) is described. BAC vector sequences were inserted into the thymidine kinase gene of HSV-2 by homologous recombination. DNA from cells infected with the resulting recombinant virus was transformed into E. coli, and colonies containing the HSV-2 BAC (HSV2-BAC) were isolated and analyzed for the expected genotype. HSV2-BAC DNA was infectious when transfected back into mammalian cells and the resulting virus was thymidine kinase negative. When used to immunize mice, the HSV2-BAC DNA elicited a strong HSV-2 specific antibody response that was equal to or greater than live virus immunization. Further, HSV2-BAC immunization was protective when animals were challenged with a lethal dose of virus. The utility of the HSV2-BAC for construction of recombinant virus genomes was demonstrated by elimination of the HSV-2 glycoprotein D (gD) gene. A recombinant HSV-2 BAC with the gD gene deleted was isolated and shown to be incapable of producing infectious virus following transfection unless an HSV gD gene was expressed in a complementing cell line. Immunization of mice with the HSV2 gD-BAC also elicited an HSV-2 specific antibody response and was protective. The results demonstrate the feasibility of DNA immunization with HSV-2 bacterial artificial chromosomes for replicating and nonreplicating candidate HSV-2 vaccines, as well as the utility of BAC technology for construction and maintenance of novel HSV-2 vaccines. The results further suggest that such technology will be a powerful tool for dissecting the immune response to HSV-2. Elsevier Inc. 2004-01-05 2003-12-02 /pmc/articles/PMC7173264/ /pubmed/14972567 http://dx.doi.org/10.1016/j.virol.2003.09.033 Text en Copyright © 2003 Elsevier Inc. All rights reserved. Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.
spellingShingle Article
Meseda, Clement A.
Schmeisser, Falko
Pedersen, Robin
Woerner, Amy
Weir, Jerry P.
DNA immunization with a herpes simplex virus 2 bacterial artificial chromosome()
title DNA immunization with a herpes simplex virus 2 bacterial artificial chromosome()
title_full DNA immunization with a herpes simplex virus 2 bacterial artificial chromosome()
title_fullStr DNA immunization with a herpes simplex virus 2 bacterial artificial chromosome()
title_full_unstemmed DNA immunization with a herpes simplex virus 2 bacterial artificial chromosome()
title_short DNA immunization with a herpes simplex virus 2 bacterial artificial chromosome()
title_sort dna immunization with a herpes simplex virus 2 bacterial artificial chromosome()
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7173264/
https://www.ncbi.nlm.nih.gov/pubmed/14972567
http://dx.doi.org/10.1016/j.virol.2003.09.033
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