AEM
Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by MacConaill, L. E.
Right arrow Articles by van Sinderen, D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by MacConaill, L. E.
Right arrow Articles by van Sinderen, D.
Agricola
Right arrow Articles by MacConaill, L. E.
Right arrow Articles by van Sinderen, D.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, December 2003, p. 6994-7001, Vol. 69, No. 12
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.12.6994-7001.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Investigation of Protein Export in Bifidobacterium breve UCC2003

Laura E. MacConaill,1,2 Gerald F. Fitzgerald,1,2,3 and Douwe van Sinderen1,2*

Alimentary Pharmabiotic Centre,1 Department of Microbiology,2 Department of Food and Nutritional Sciences, National University of Ireland, Cork, Cork, Ireland3

Received 21 May 2003/ Accepted 8 September 2003

The molecular interactions between the bifidobacterial cell and its natural environment, namely, the gastrointestinal tract of its host, are particularly important in understanding the presumed positive effects of Bifidobacterium on the health status of the host. In this study an export-specific reporter system, designed for use in gram-positive organisms and based on the use of the staphylococcal nuclease (Nuc) as a reporter, was employed to identify exported proteins in Bifidobacterium breve UCC2003. A B. breve genomic library of translational fusions to the Nuc-encoding gene devoid of its own export signal was established in the shuttle vector pFUN (I. Poquet, S. D. Ehrlich, and A. Gruss, J. Bacteriol. 180:1904-1912, 1998) and screened for bifidobacterial export signals. Sequence analysis of the fusion proteins obtained that displayed a nuclease-producing phenotype in both Lactococcus lactis and B. breve predicted the presence of a classical signal peptide and/or single or multiple transmembrane domains, thus indicating that some of the export signals in B. breve are comparable to those used in L. lactis. Cell fractionation studies, zymograms, nuclease assays, and Western blotting were employed to confirm the function of the predicted signals and to determine the location and activity of the exported fusion proteins in B. breve and/or L. lactis.

* Corresponding author. Mailing address: Department of Microbiology, National University of Ireland, Cork, Western Road, Cork, Ireland. Phone: 353 21 4901365. Fax: 353 21 4903101. E-mail: d.vansinderen{at}ucc.ie.

Applied and Environmental Microbiology, December 2003, p. 6994-7001, Vol. 69, No. 12
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.12.6994-7001.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:



Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
J. Bacteriol. Microbiol. Mol. Biol. Rev. Eukaryot. Cell All ASM Journals

Copyright © 2003 by the American Society for Microbiology. All rights reserved.