Permeabilization of the mycobacterial envelope for protein cytolocalization studies by immunofluorescence microscopy.
BACKGROUND
The establishment of the cellular localization of proteins in M. tuberculosis will provide of valuable information for the identification of new drug/vaccine/diagnostic targets. Cytolocalization by inmunofluorescence microscopy has been limited in mycobacteria because to difficulties in effectively permeabilize it.


RESULTS
A treatment combining lysozyme with triton X-100 was found to be an effective permeabilization method of the mycobacterial envelope.


CONCLUSIONS
A rapid and simple permeabilization protocol has been successfully assessed in pure cultures of both Mycobacterium smegmatis and Mycobacterium tuberculosis H37Rv. This method can be successful used in the cytolocalization of proteins by immunolabeling.
Cimino
Mena
M
Departamento de Biología Estructural, Instituto Venezolano de Investigaciones Científicas, Altos de Pipe, Estado Miranda, Venezuela. mcimino@ivic.ve
Alamo
Lorenzo
L
Salazar
Leiria
L
D001426:Bacterial Proteins / Q000276:immunology / Q000378:metabolism*; D002462:Cell Membrane / Q000737:chemistry / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D002463:Cell Membrane Permeability / Q000187:drug effects; D008856:Microscopy, Fluorescence / Q000379:methods*; D009113:Muramidase / Q000737:chemistry / Q000494:pharmacology; D020102:Mycobacterium smegmatis / Q000166:cytology* / Q000187:drug effects; D009169:Mycobacterium tuberculosis / Q000166:cytology* / Q000187:drug effects; D017830:Octoxynol / Q000737:chemistry / Q000494:pharmacology; D021381:Protein Transport
D001426:Bacterial Proteins; D017830:Octoxynol; D009113:Muramidase