Open Access Highly Accessed Research article

Characterisation of porin genes from Mycobacterium fortuitum and their impact on growth

Soroush Sharbati12, Kira Schramm2, Sonja Rempel2, Hwa Wang2, Ronny Andrich2, Verena Tykiel3, Ralph Kunisch2 and Astrid Lewin2*

Author Affiliations

1 Freie Universität Berlin, Institute of Veterinary Biochemistry, Oertzenweg 19b, 14163 Berlin, Germany

2 Robert Koch-Institute, Nordufer 20, 13533 Berlin, Germany

3 Deutsches Institut für Bautechnik (DIBt) Section II 4 – Health and Environmental Protection, Kolonnenstr. 30 L, 10829 Berlin, Germany

For all author emails, please log on.

BMC Microbiology 2009, 9:31  doi:10.1186/1471-2180-9-31

Published: 9 February 2009

Additional files

Additional file 1:

Growth rate of the M. fortuitum strains 10851/03, 10860/03 and DSM 46621. Logarithmic display of the growth curves shown in Figure 1. The growth rate of the strains was measured by quantification of the ATP-content [displayed as relative light units (RLU)] in broth cultures.

Format: PPT Size: 50KB Download file

This file can be viewed with: Microsoft PowerPoint Viewer

Open Data

Additional file 2:

Detection of the PorM spot on the 2D-PAGE by Western Blot analysis. Detection was performed using the porin-specific antiserum pAK MspA#813 on the blotted 2D-PAGE shown in Figure 5A. Only one protein spot was identified possessing an apparent molecular mass of approximately 120 kDa and an apparent pI of about 4. The arrow indicates the identified spot.

Format: PPT Size: 319KB Download file

This file can be viewed with: Microsoft PowerPoint Viewer

Open Data

Additional file 3:

Western Blot analysis of PorMs in M. fortuitum. Porin expression in members of the M. fortuitum-group was studied by Western blotting. 10–30 μg of protein extracted with nOPOE was separated by 1D-SDS-PAGE and detected by the antiserum pAK MspA#813. Lanes 1–4: 1, M. smegmatis SMR5 (10 μg); 2, M. fortuitum DSM 466211 (30 μg); 3, M. fortuitum 10851/03 (30 μg); 4, M. fortuitum 10860/03 (30 μg).

Format: PPT Size: 161KB Download file

This file can be viewed with: Microsoft PowerPoint Viewer

Open Data

Additional file 4:

Detection of PorMs on the surface of M. fortuitum. Detection was performed using the porin-specific antiserum pAK MspA#813 in quantitative microwell immunoassays. Each column represents the mean (± SD) of 8 measurements. Asterisks indicate significant differences between the samples, which were treated with pAK MspA#813 and backgrounds according to the paired Student's t-test (P < 0.001).

Format: PPT Size: 85KB Download file

This file can be viewed with: Microsoft PowerPoint Viewer

Open Data

Additional file 5:

Knock-down of porins in M. fortuitum 10860/03 by means of anti-sense technology using the plasmid pSRr106. The amount of porM1/porM2 mRNA was quantified by means of qRT-PCR and was normalised with 16S rRNA. Compared to the reference strain M. fortuitum 10860/03 (pSHKLx1) the amount of porM mRNA in the down-regulated strain 10860/03 (pSRr106) was reduced by about 75%.

Format: PPT Size: 52KB Download file

This file can be viewed with: Microsoft PowerPoint Viewer

Open Data