Generation of affinity-tagged fluoromycobacteriophages by mixed assembly of phage capsids
Addition of affinity tags to bacteriophage particles facilitates a variety of applications, including vaccine construction and diagnosis of bacterial infections. Addition of tags to phage capsids is desirable, as modification of the tails can lead to poor adsorption and loss of infectivity. Although...
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| Formato: | Artículo publishedVersion |
| Lenguaje: | Inglés |
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2013
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| Acceso en línea: | http://hdl.handle.net/20.500.12110/paper_00992240_v79_n18_p5608_Piuri |
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paperaa:paper_00992240_v79_n18_p5608_Piuri2023-06-12T16:46:25Z Generation of affinity-tagged fluoromycobacteriophages by mixed assembly of phage capsids Appl. Environ. Microbiol. 2013;79(18):5608-5615 Piuri, M. Rondón, L. Urdániz, E. Hatfull, G.F. Affinity tags Bacterial infections Mixed assemblies Mycobacteriophage Wild types Bacteria Bacteriophages Proteins adsorption bacteriophage bacterium gene expression growth rate homogeneity identification method infectivity recombination capsid protein hybrid protein article fluorescence genetics isolation and purification metabolism methodology microbiological examination mycobacteriophage Mycobacterium physiology staining virology virus assembly virus capsid Bacteriological Techniques Capsid Capsid Proteins Fluorescence Mycobacteriophages Mycobacterium Recombinant Fusion Proteins Staining and Labeling Virus Assembly Addition of affinity tags to bacteriophage particles facilitates a variety of applications, including vaccine construction and diagnosis of bacterial infections. Addition of tags to phage capsids is desirable, as modification of the tails can lead to poor adsorption and loss of infectivity. Although tags can readily be included as fusions to head decoration proteins, many phages do not have decoration proteins as virion components. The addition of a small (10-amino-acid) Strep-tag II (STAG II) to the mycobacteriophage TM4 capsid subunit, gp9, was not tolerated as a genetically homogenous recombinant phage but could be incorporated into the head by growth of wild-type phage on a host expressing the capsid-STAG fusion. Particles with capsids composed of wild-type and STAG-tagged subunit mixtures could be grown to high titers, showed good infectivities, and could be used to isolate phage-bacterium complexes. Preparation of a STAG-labeled fluoromycobacteriophage enabled capture of bacterial complexes and identification of infected bacteria by fluorescence. © 2013, American Society for Microbiology. Fil:Piuri, M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2013 info:eu-repo/semantics/article info:ar-repo/semantics/artículo info:eu-repo/semantics/publishedVersion application/pdf eng info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_00992240_v79_n18_p5608_Piuri |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| language |
Inglés |
| orig_language_str_mv |
eng |
| topic |
Affinity tags Bacterial infections Mixed assemblies Mycobacteriophage Wild types Bacteria Bacteriophages Proteins adsorption bacteriophage bacterium gene expression growth rate homogeneity identification method infectivity recombination capsid protein hybrid protein article fluorescence genetics isolation and purification metabolism methodology microbiological examination mycobacteriophage Mycobacterium physiology staining virology virus assembly virus capsid Bacteriological Techniques Capsid Capsid Proteins Fluorescence Mycobacteriophages Mycobacterium Recombinant Fusion Proteins Staining and Labeling Virus Assembly |
| spellingShingle |
Affinity tags Bacterial infections Mixed assemblies Mycobacteriophage Wild types Bacteria Bacteriophages Proteins adsorption bacteriophage bacterium gene expression growth rate homogeneity identification method infectivity recombination capsid protein hybrid protein article fluorescence genetics isolation and purification metabolism methodology microbiological examination mycobacteriophage Mycobacterium physiology staining virology virus assembly virus capsid Bacteriological Techniques Capsid Capsid Proteins Fluorescence Mycobacteriophages Mycobacterium Recombinant Fusion Proteins Staining and Labeling Virus Assembly Piuri, M. Rondón, L. Urdániz, E. Hatfull, G.F. Generation of affinity-tagged fluoromycobacteriophages by mixed assembly of phage capsids |
| topic_facet |
Affinity tags Bacterial infections Mixed assemblies Mycobacteriophage Wild types Bacteria Bacteriophages Proteins adsorption bacteriophage bacterium gene expression growth rate homogeneity identification method infectivity recombination capsid protein hybrid protein article fluorescence genetics isolation and purification metabolism methodology microbiological examination mycobacteriophage Mycobacterium physiology staining virology virus assembly virus capsid Bacteriological Techniques Capsid Capsid Proteins Fluorescence Mycobacteriophages Mycobacterium Recombinant Fusion Proteins Staining and Labeling Virus Assembly |
| description |
Addition of affinity tags to bacteriophage particles facilitates a variety of applications, including vaccine construction and diagnosis of bacterial infections. Addition of tags to phage capsids is desirable, as modification of the tails can lead to poor adsorption and loss of infectivity. Although tags can readily be included as fusions to head decoration proteins, many phages do not have decoration proteins as virion components. The addition of a small (10-amino-acid) Strep-tag II (STAG II) to the mycobacteriophage TM4 capsid subunit, gp9, was not tolerated as a genetically homogenous recombinant phage but could be incorporated into the head by growth of wild-type phage on a host expressing the capsid-STAG fusion. Particles with capsids composed of wild-type and STAG-tagged subunit mixtures could be grown to high titers, showed good infectivities, and could be used to isolate phage-bacterium complexes. Preparation of a STAG-labeled fluoromycobacteriophage enabled capture of bacterial complexes and identification of infected bacteria by fluorescence. © 2013, American Society for Microbiology. |
| format |
Artículo Artículo publishedVersion |
| author |
Piuri, M. Rondón, L. Urdániz, E. Hatfull, G.F. |
| author_facet |
Piuri, M. Rondón, L. Urdániz, E. Hatfull, G.F. |
| author_sort |
Piuri, M. |
| title |
Generation of affinity-tagged fluoromycobacteriophages by mixed assembly of phage capsids |
| title_short |
Generation of affinity-tagged fluoromycobacteriophages by mixed assembly of phage capsids |
| title_full |
Generation of affinity-tagged fluoromycobacteriophages by mixed assembly of phage capsids |
| title_fullStr |
Generation of affinity-tagged fluoromycobacteriophages by mixed assembly of phage capsids |
| title_full_unstemmed |
Generation of affinity-tagged fluoromycobacteriophages by mixed assembly of phage capsids |
| title_sort |
generation of affinity-tagged fluoromycobacteriophages by mixed assembly of phage capsids |
| publishDate |
2013 |
| url |
http://hdl.handle.net/20.500.12110/paper_00992240_v79_n18_p5608_Piuri |
| work_keys_str_mv |
AT piurim generationofaffinitytaggedfluoromycobacteriophagesbymixedassemblyofphagecapsids AT rondonl generationofaffinitytaggedfluoromycobacteriophagesbymixedassemblyofphagecapsids AT urdanize generationofaffinitytaggedfluoromycobacteriophagesbymixedassemblyofphagecapsids AT hatfullgf generationofaffinitytaggedfluoromycobacteriophagesbymixedassemblyofphagecapsids |
| _version_ |
1769810177254490112 |