Mycobacterial proteome extraction: comparison of disruption methods.

Abstract:

:Mycobacterium avium subsp. paratuberculosis has long been recognized as the causative agent of Johne's disease, a chronic inflammatory intestinal disease of sheep, cattle and other ruminants. Mycobacterial cells are extremely hardy, and proteomic analyses require the use of harsh conditions to effect their disruption. We compared the effectiveness of bead beating and sonication as cell lysis methods for the extraction of the proteomes of Mycobacterium avium subsp. avium and Mycobacterium avium subsp. paratuberculosis. Broad and narrow range two-dimensional gel electrophoresis was used to compare the numbers of silver stained protein spots that were observed in mycobacterial lysates. Despite differences in the yield of total protein from either species, and at different ages, the two methods appeared to give similar representations of the mycobacterial proteomes analyzed. Bead beating therefore represents a rapid and effective method of extracting the proteomes of mycobacterial species without the risks associated with an open tube sonication procedure.

journal_name

Proteomics

journal_title

Proteomics

authors

Lanigan MD,Vaughan JA,Shiell BJ,Beddome GJ,Michalski WP

doi

10.1002/pmic.200300672

subject

Has Abstract

pub_date

2004-04-01 00:00:00

pages

1094-100

issue

4

eissn

1615-9853

issn

1615-9861

journal_volume

4

pub_type

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