Generation of affinity-tagged fluoromycobacteriophages by mixed assembly of phage capsids

PIURI MARIANA; LILIANA RONDON; ESTEFANIA URDANIZ; GRAHAM F HATFULL

APPLIED AND ENVIRONMENTAL MICROBIOLOGY

AMER SOC MICROBIOLOGY

Lugar: Washington; Año: 2013 vol. 79 p. 5608 - 5615

0099-2240

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 aa) Strep II tag (STAG) 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 titer, show good infectivity, and can be used to isolate phage-bacterial complexes.  Preparation of a STAG-labeled fluoromycobacteriophage enabled capture of bacterial complexes and identification of infected bacteria by fluorescence.