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AuthorSun, Zhongkedc.contributor.author
Date of accession2016-03-15T10:40:35Zdc.date.accessioned
Available in OPARU since2016-03-15T10:40:35Zdc.date.available
Year of creation2014dc.date.created
AbstractBased on their beneficial properties and their generally being recognized as safe status, bifidobacteria are used as probiotics in food and pharmaceutical industries. In the future, recombinant strains with enhanced and/or new beneficial properties may become an option for alternative or supplementary treatment of various diseases including inflammatory bowel disease or cancer. This thesis established an efficient system for production of secreted proteins with therapeutic potential using recombinant bifidobacteria and composed by following work: 1. Four putative promoter sequences were analyzed using a glucuronidase reporter system in B. bifidum S17. This revealed that the DNA sequences upstream of the gap gene yielded highest levels of GusA reporter activity amongst the promoters tested. 2. The transcription start site of the gap promoter was determined to be a thymidine residue 62 bp upstream of the translational start site. Using bioinformatics comparison, the -10, -35 and ribosome-binding site of Pgap were identified. The Pgap core regions are highly conserved in all five Bifidobacterium sp. analyzed indicating that Pgap of B. bifidum S17 is functional across these species. 3. Using the E. coli appA gene and the predicted signal peptide of a sialidase (BBIF_1734), a reporter vector for analysis of signal peptides in B. bifidum S17 was generated. This system was used to analyze the efficacy of other seven predicted signal peptides to mediate protein secretion. 4. The most efficient signal peptide were fused to a codon adapted gene encoding for a nitroreductase, and cloned under control of Pgap. The resulting vector was transformed into B. bifidum S17 and this recombinant strain exhibited high nitroreductase activity in culture supernatants. The engineered strain can efficiently convert the prodrug CB1954 to cytotoxic drug.dc.description.abstract
Languageendc.language.iso
PublisherUniversität Ulmdc.publisher
LicenseStandarddc.rights
Link to license texthttps://oparu.uni-ulm.de/xmlui/license_v3dc.rights.uri
KeywordBacterial tumor therapydc.subject
KeywordReporterdc.subject
Dewey Decimal GroupDDC 570 / Life sciencesdc.subject.ddc
LCSHGene expression Prostatadc.subject.lcsh
LCSHSecretiondc.subject.lcsh
TitleDevelopment of gene expression systems in Bifidobacterium bifidum S17 and their application for tumor therapydc.title
Resource typeDissertationdc.type
DOIhttp://dx.doi.org/10.18725/OPARU-3304dc.identifier.doi
PPN80691257Xdc.identifier.ppn
URNhttp://nbn-resolving.de/urn:nbn:de:bsz:289-vts-92777dc.identifier.urn
GNDBifidobacteriumdc.subject.gnd
GNDGenexpressiondc.subject.gnd
FacultyFakultät für Naturwissenschaftenuulm.affiliationGeneral
Date of activation2014-11-19T10:59:27Zuulm.freischaltungVTS
Peer reviewneinuulm.peerReview
Shelfmark print versionW: W-H 13.869uulm.shelfmark
DCMI TypeTextuulm.typeDCMI
VTS ID9277uulm.vtsID
CategoryPublikationenuulm.category
Bibliographyuulmuulm.bibliographie


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