Molecular analysis of the acinetobacter baumannii biofilm-associated protein
Acinetobacter baumannii is a multidrug-resistant pathogen associated with hospital outbreaks of infection across the globe, particularly in the intensive care unit. The ability of A. baumannii to survive in the hospital environment for long periods is linked to antibiotic resistance and its capacity...
Saved in:
Main Authors: | , , , , , , , , , , , |
---|---|
Other Authors: | |
Format: | Article |
Language: | English |
Published: |
2015
|
Subjects: | |
Online Access: | https://hdl.handle.net/10356/103048 http://hdl.handle.net/10220/25760 |
Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
Institution: | Nanyang Technological University |
Language: | English |
id |
sg-ntu-dr.10356-103048 |
---|---|
record_format |
dspace |
spelling |
sg-ntu-dr.10356-1030482022-02-16T16:28:54Z Molecular analysis of the acinetobacter baumannii biofilm-associated protein Sidjabat, Hanna E. Paterson, David L. Nimmo, Graeme R. Lipman, Jeffrey Schembri, Mark A. Goh, Hwee Mian Sharon Beatson, Scott A. Totsika, Makrina Moriel, Danilo G. Phan, Minh-Duy Szubert, Jan Runnegar, Naomi Singapore Centre for Environmental Life Sciences Engineering DRNTU::Science::Biological sciences::Microbiology Acinetobacter baumannii is a multidrug-resistant pathogen associated with hospital outbreaks of infection across the globe, particularly in the intensive care unit. The ability of A. baumannii to survive in the hospital environment for long periods is linked to antibiotic resistance and its capacity to form biofilms. Here we studied the prevalence, expression, and function of the A. baumannii biofilm-associated protein (Bap) in 24 carbapenem-resistant A. baumannii ST92 strains isolated from a single institution over a 10-year period. The bap gene was highly prevalent, with 22/24 strains being positive for bap by PCR. Partial sequencing of bap was performed on the index case strain MS1968 and revealed it to be a large and highly repetitive gene approximately 16 kb in size. Phylogenetic analysis employing a 1,948-amino-acid region corresponding to the C terminus of Bap showed that BapMS1968 clusters with Bap sequences from clonal complex 2 (CC2) strains ACICU, TCDC-AB0715, and 1656-2 and is distinct from Bap in CC1 strains. By using overlapping PCR, the bapMS1968 gene was cloned, and its expression in a recombinant Escherichia coli strain resulted in increased biofilm formation. A Bap-specific antibody was generated, and Western blot analysis showed that the majority of A. baumannii strains expressed an ∼200-kDa Bap protein. Further analysis of three Bap-positive A. baumannii strains demonstrated that Bap is expressed at the cell surface and is associated with biofilm formation. Finally, biofilm formation by these Bap-positive strains could be inhibited by affinity-purified Bap antibodies, demonstrating the direct contribution of Bap to biofilm growth by A. baumannii clinical isolates. Published version 2015-06-04T08:32:33Z 2019-12-06T21:04:25Z 2015-06-04T08:32:33Z 2019-12-06T21:04:25Z 2013 2013 Journal Article Goh, H. M. S., Beatson, S. A., Totsika, M., Moriel, D. G., Phan, M.-D., Szubert, J., et al. (2013). Molecular analysis of the acinetobacter baumannii biofilm-associated protein. Applied and environmental microbiology, 79(21), 6535-6543. 0099-2240 https://hdl.handle.net/10356/103048 http://hdl.handle.net/10220/25760 10.1128/AEM.01402-13 23956398 en Applied and environmental microbiology © 2013 American Society for Microbiology (ASM). This paper was published in Applied and Environmental Microbiology and is made available as an electronic reprint (preprint) with permission of American Society for Microbiology (ASM). The paper can be found at the following official DOI: [http://dx.doi.org/10.1128/AEM.01402-13]. One print or electronic copy may be made for personal use only. Systematic or multiple reproduction, distribution to multiple locations via electronic or other means, duplication of any material in this paper for a fee or for commercial purposes, or modification of the content of the paper is prohibited and is subject to penalties under law. application/pdf |
institution |
Nanyang Technological University |
building |
NTU Library |
continent |
Asia |
country |
Singapore Singapore |
content_provider |
NTU Library |
collection |
DR-NTU |
language |
English |
topic |
DRNTU::Science::Biological sciences::Microbiology |
spellingShingle |
DRNTU::Science::Biological sciences::Microbiology Sidjabat, Hanna E. Paterson, David L. Nimmo, Graeme R. Lipman, Jeffrey Schembri, Mark A. Goh, Hwee Mian Sharon Beatson, Scott A. Totsika, Makrina Moriel, Danilo G. Phan, Minh-Duy Szubert, Jan Runnegar, Naomi Molecular analysis of the acinetobacter baumannii biofilm-associated protein |
description |
Acinetobacter baumannii is a multidrug-resistant pathogen associated with hospital outbreaks of infection across the globe, particularly in the intensive care unit. The ability of A. baumannii to survive in the hospital environment for long periods is linked to antibiotic resistance and its capacity to form biofilms. Here we studied the prevalence, expression, and function of the A. baumannii biofilm-associated protein (Bap) in 24 carbapenem-resistant A. baumannii ST92 strains isolated from a single institution over a 10-year period. The bap gene was highly prevalent, with 22/24 strains being positive for bap by PCR. Partial sequencing of bap was performed on the index case strain MS1968 and revealed it to be a large and highly repetitive gene approximately 16 kb in size. Phylogenetic analysis employing a 1,948-amino-acid region corresponding to the C terminus of Bap showed that BapMS1968 clusters with Bap sequences from clonal complex 2 (CC2) strains ACICU, TCDC-AB0715, and 1656-2 and is distinct from Bap in CC1 strains. By using overlapping PCR, the bapMS1968 gene was cloned, and its expression in a recombinant Escherichia coli strain resulted in increased biofilm formation. A Bap-specific antibody was generated, and Western blot analysis showed that the majority of A. baumannii strains expressed an ∼200-kDa Bap protein. Further analysis of three Bap-positive A. baumannii strains demonstrated that Bap is expressed at the cell surface and is associated with biofilm formation. Finally, biofilm formation by these Bap-positive strains could be inhibited by affinity-purified Bap antibodies, demonstrating the direct contribution of Bap to biofilm growth by A. baumannii clinical isolates. |
author2 |
Singapore Centre for Environmental Life Sciences Engineering |
author_facet |
Singapore Centre for Environmental Life Sciences Engineering Sidjabat, Hanna E. Paterson, David L. Nimmo, Graeme R. Lipman, Jeffrey Schembri, Mark A. Goh, Hwee Mian Sharon Beatson, Scott A. Totsika, Makrina Moriel, Danilo G. Phan, Minh-Duy Szubert, Jan Runnegar, Naomi |
format |
Article |
author |
Sidjabat, Hanna E. Paterson, David L. Nimmo, Graeme R. Lipman, Jeffrey Schembri, Mark A. Goh, Hwee Mian Sharon Beatson, Scott A. Totsika, Makrina Moriel, Danilo G. Phan, Minh-Duy Szubert, Jan Runnegar, Naomi |
author_sort |
Sidjabat, Hanna E. |
title |
Molecular analysis of the acinetobacter baumannii biofilm-associated protein |
title_short |
Molecular analysis of the acinetobacter baumannii biofilm-associated protein |
title_full |
Molecular analysis of the acinetobacter baumannii biofilm-associated protein |
title_fullStr |
Molecular analysis of the acinetobacter baumannii biofilm-associated protein |
title_full_unstemmed |
Molecular analysis of the acinetobacter baumannii biofilm-associated protein |
title_sort |
molecular analysis of the acinetobacter baumannii biofilm-associated protein |
publishDate |
2015 |
url |
https://hdl.handle.net/10356/103048 http://hdl.handle.net/10220/25760 |
_version_ |
1725985653178499072 |