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A general insert label for peptide display on chimeric filamentous bacteriophages

The foreign insert intended to be displayed via recombinant phage proteins can have a negative effect on protein expression and phage assembly. A typical example is the case of display of peptides longer than 6 amino acid residues on the major coat protein, protein VIII of the filamentous bacterioph...

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Detalles Bibliográficos
Autores principales: Kaplan, Gilad, Gershoni, Jonathan M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier Inc. 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7094602/
https://www.ncbi.nlm.nih.gov/pubmed/21945353
http://dx.doi.org/10.1016/j.ab.2011.08.050
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author Kaplan, Gilad
Gershoni, Jonathan M.
author_facet Kaplan, Gilad
Gershoni, Jonathan M.
author_sort Kaplan, Gilad
collection PubMed
description The foreign insert intended to be displayed via recombinant phage proteins can have a negative effect on protein expression and phage assembly. A typical example is the case of display of peptides longer than 6 amino acid residues on the major coat protein, protein VIII of the filamentous bacteriophages M13 and fd. A solution to this problem has been the use of “two-gene systems” generating chimeric phages that concomitantly express wild-type protein VIII along with recombinant protein VIII. Although the two-gene systems are much more permissive in regard to insert length and composition, some cases can still adversely affect phage assembly. Although these phages genotypically contain the desired DNA of the insert, they appear to be phenotypically wild type. To avoid false-negative results when using chimeric phages in binding studies, it is necessary to confirm that the observed lack of phage recognition is not due to faulty assembly and display of the intended insert. Here we describe a strategy for generating antibodies that specifically recognize recombinant protein VIII regardless of the nature of its foreign insert. These antibodies can be used as a general monitor of the display of recombinant protein VIII into phage particles.
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spelling pubmed-70946022020-03-25 A general insert label for peptide display on chimeric filamentous bacteriophages Kaplan, Gilad Gershoni, Jonathan M. Anal Biochem Article The foreign insert intended to be displayed via recombinant phage proteins can have a negative effect on protein expression and phage assembly. A typical example is the case of display of peptides longer than 6 amino acid residues on the major coat protein, protein VIII of the filamentous bacteriophages M13 and fd. A solution to this problem has been the use of “two-gene systems” generating chimeric phages that concomitantly express wild-type protein VIII along with recombinant protein VIII. Although the two-gene systems are much more permissive in regard to insert length and composition, some cases can still adversely affect phage assembly. Although these phages genotypically contain the desired DNA of the insert, they appear to be phenotypically wild type. To avoid false-negative results when using chimeric phages in binding studies, it is necessary to confirm that the observed lack of phage recognition is not due to faulty assembly and display of the intended insert. Here we describe a strategy for generating antibodies that specifically recognize recombinant protein VIII regardless of the nature of its foreign insert. These antibodies can be used as a general monitor of the display of recombinant protein VIII into phage particles. Elsevier Inc. 2012-01-01 2011-09-08 /pmc/articles/PMC7094602/ /pubmed/21945353 http://dx.doi.org/10.1016/j.ab.2011.08.050 Text en Copyright © 2011 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
Kaplan, Gilad
Gershoni, Jonathan M.
A general insert label for peptide display on chimeric filamentous bacteriophages
title A general insert label for peptide display on chimeric filamentous bacteriophages
title_full A general insert label for peptide display on chimeric filamentous bacteriophages
title_fullStr A general insert label for peptide display on chimeric filamentous bacteriophages
title_full_unstemmed A general insert label for peptide display on chimeric filamentous bacteriophages
title_short A general insert label for peptide display on chimeric filamentous bacteriophages
title_sort general insert label for peptide display on chimeric filamentous bacteriophages
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7094602/
https://www.ncbi.nlm.nih.gov/pubmed/21945353
http://dx.doi.org/10.1016/j.ab.2011.08.050
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