Permeabilization of the mycobacterial envelope for protein cytolocalization studies by immunofluorescence microscopy-Reference-Cited by-同舟云学术

Permeabilization of the mycobacterial envelope for protein cytolocalization studies by immunofluorescence microscopy

Published:2006-04-18 Issue:1 Volume:6 Page:
ISSN:1471-2180
Container-title:BMC Microbiology
language:en
Short-container-title:BMC Microbiol

Author:

Cimino Mena,Alamo Lorenzo,Salazar Leiria

Abstract

Abstract 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. Conclusion 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.

Publisher

Springer Science and Business Media LLC

Subject

Microbiology (medical),Microbiology

Link

https://link.springer.com/content/pdf/10.1186/1471-2180-6-35.pdf

Reference14 articles.

1. Allan VJ: Basic immunofluorescence. protein localization by fluorescent microscopy. A practical approach. Edited by: Allan VJ. 2000, Oxford University Press. USA

2. Carr E, Kathryn E, Soddell J, Seviour R: Improved permeabilization protocols for fluorescence in situ hybridization (FISH) of mycolic-acid-containing bacteria found in foams. J Microbiol Methods. 2005, 61: 47-54. 10.1016/j.mimet.2004.10.023.

3. Cole ST, Brosh R, Parkhill J, Garnier T, Churcher C, Harris D, Gordon SV, Eiglmeier K, Gas S, Barry CE, Tekaia F, Badcock K, Basham D, Brown D, Chillingworth T, Connor R, Davies R, Devlin K, Feltwell T, Gentles S, Hamlin N, Holroyd S, Hornsby T, Jagels K, Krogh A, McLean J, Moule S, Murphy L, Oliver K, Osborne J, Quail MA, Rajandream MA, Rogers J, Rutter S, Seeger K, Skelton J, Squares R, Squares S, Sulston JE, Taylor K, Whitehead S, Barrell BG: Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequence. Nature. 1998, 393: 537-544. 10.1038/31159.

4. Delogu G, Pusceddu C, Bua A, Fadda G, Brennan MJ, Zanetti S: Rv1818c-encoded PE_PGRS protein of Mycobacterium tuberculosis is surface exposed and influences bacterial cell structure. Mol Microbiol. 2004, 52: 725-33. 10.1111/j.1365-2958.2004.04007.x.

5. Devenport RJ, Curtis TP, Goodfellow M, Stainsby FM, Bingley M: Quantitative use of fluorescent in situ hybridization to examine relationships between mycolic acid-containing actinomycetes and foaming in activated sludge plants. Appl Environ Microbiol. 2000, 66: 1158-1166. 10.1128/AEM.66.3.1158-1166.2000.

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