IRIS publication 43337709
Investigation of protein export in Bifidobacterium breve UCC2003
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TY - JOUR - MacConaill, LE,Fitzgerald, GF,van Sinderen, D - 2003 - January - Applied and Environmental Microbiology - Investigation of protein export in Bifidobacterium breve UCC2003 - Validated - () - GRAM-POSITIVE BACTERIA BACILLUS-SUBTILIS ESCHERICHIA-COLI STAPHYLOCOCCAL NUCLEASE LACTOCOCCUS-LACTIS SIGNAL PEPTIDE STREPTOCOCCUS-LACTIS MOLECULAR-CLONING MEMBRANE-PROTEINS GENOME SEQUENCE - 69 - 6994 - 7001 - 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. - DOI 10.1128/AEM.69.12.6994-7001.2003 DA - 2003/01 ER -
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@article{V43337709, = {MacConaill, LE and Fitzgerald, GF and van Sinderen, D }, = {2003}, = {January}, = {Applied and Environmental Microbiology}, = {Investigation of protein export in Bifidobacterium breve UCC2003}, = {Validated}, = {()}, = {GRAM-POSITIVE BACTERIA BACILLUS-SUBTILIS ESCHERICHIA-COLI STAPHYLOCOCCAL NUCLEASE LACTOCOCCUS-LACTIS SIGNAL PEPTIDE STREPTOCOCCUS-LACTIS MOLECULAR-CLONING MEMBRANE-PROTEINS GENOME SEQUENCE}, = {69}, pages = {6994--7001}, = {{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.}}, = {DOI 10.1128/AEM.69.12.6994-7001.2003}, source = {IRIS} }
Data as stored in IRIS
AUTHORS | MacConaill, LE,Fitzgerald, GF,van Sinderen, D | ||
YEAR | 2003 | ||
MONTH | January | ||
JOURNAL_CODE | Applied and Environmental Microbiology | ||
TITLE | Investigation of protein export in Bifidobacterium breve UCC2003 | ||
STATUS | Validated | ||
TIMES_CITED | () | ||
SEARCH_KEYWORD | GRAM-POSITIVE BACTERIA BACILLUS-SUBTILIS ESCHERICHIA-COLI STAPHYLOCOCCAL NUCLEASE LACTOCOCCUS-LACTIS SIGNAL PEPTIDE STREPTOCOCCUS-LACTIS MOLECULAR-CLONING MEMBRANE-PROTEINS GENOME SEQUENCE | ||
VOLUME | 69 | ||
ISSUE | |||
START_PAGE | 6994 | ||
END_PAGE | 7001 | ||
ABSTRACT | 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. | ||
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DOI_LINK | DOI 10.1128/AEM.69.12.6994-7001.2003 | ||
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