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Abstract:

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.

Registro:

Documento: Artículo
Título:Generation of affinity-tagged fluoromycobacteriophages by mixed assembly of phage capsids
Autor:Piuri, M.; Rondón, L.; Urdániz, E.; Hatfull, G.F.
Filiación:Department of Biological Sciences, University of Pittsburgh, Pittsburgh, PA, United States
Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, IQUIBICEN-CONICET, Buenos Aires, Argentina
Palabras clave: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
Año:2013
Volumen:79
Número:18
Página de inicio:5608
Página de fin:5615
DOI: http://dx.doi.org/10.1128/AEM.01016-13
Título revista:Applied and Environmental Microbiology
Título revista abreviado:Appl. Environ. Microbiol.
ISSN:00992240
CODEN:AEMID
PDF:https://bibliotecadigital.exactas.uba.ar/download/paper/paper_00992240_v79_n18_p5608_Piuri.pdf
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00992240_v79_n18_p5608_Piuri

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Citas:

---------- APA ----------
Piuri, M., Rondón, L., Urdániz, E. & Hatfull, G.F. (2013) . Generation of affinity-tagged fluoromycobacteriophages by mixed assembly of phage capsids. Applied and Environmental Microbiology, 79(18), 5608-5615.
http://dx.doi.org/10.1128/AEM.01016-13
---------- CHICAGO ----------
Piuri, M., Rondón, L., Urdániz, E., Hatfull, G.F. "Generation of affinity-tagged fluoromycobacteriophages by mixed assembly of phage capsids" . Applied and Environmental Microbiology 79, no. 18 (2013) : 5608-5615.
http://dx.doi.org/10.1128/AEM.01016-13
---------- MLA ----------
Piuri, M., Rondón, L., Urdániz, E., Hatfull, G.F. "Generation of affinity-tagged fluoromycobacteriophages by mixed assembly of phage capsids" . Applied and Environmental Microbiology, vol. 79, no. 18, 2013, pp. 5608-5615.
http://dx.doi.org/10.1128/AEM.01016-13
---------- VANCOUVER ----------
Piuri, M., Rondón, L., Urdániz, E., Hatfull, G.F. Generation of affinity-tagged fluoromycobacteriophages by mixed assembly of phage capsids. Appl. Environ. Microbiol. 2013;79(18):5608-5615.
http://dx.doi.org/10.1128/AEM.01016-13