• English
    • Deutsch
  • English 
    • English
    • Deutsch
  • Login
View Item 
  •   Home
  • Universität Ulm
  • Publikationen
  • View Item
  •   Home
  • Universität Ulm
  • Publikationen
  • View Item
JavaScript is disabled for your browser. Some features of this site may not work without it.

Development of gene expression systems in Bifidobacterium bifidum S17 and their application for tumor therapy

Thumbnail
vts_9277_13937.pdf (1.705Mb)
103 S.
Veröffentlichung
2014-11-19
Authors
Sun, Zhongke
Dissertation


Faculties
Fakultät für Naturwissenschaften
Abstract
Based 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.
Date created
2014
Subject headings
[GND]: Bifidobacterium | Genexpression
[LCSH]: Gene expression Prostata | Secretion
[Free subject headings]: Bacterial tumor therapy | Reporter
[DDC subject group]: DDC 570 / Life sciences
License
Standard
https://oparu.uni-ulm.de/xmlui/license_v3

Metadata
Show full item record

DOI & citation

Please use this identifier to cite or link to this item: http://dx.doi.org/10.18725/OPARU-3304

Sun, Zhongke (2014): Development of gene expression systems in Bifidobacterium bifidum S17 and their application for tumor therapy. Open Access Repositorium der Universität Ulm und Technischen Hochschule Ulm. Dissertation. http://dx.doi.org/10.18725/OPARU-3304
Citation formatter >



Policy | kiz service OPARU | Contact Us
Impressum | Privacy statement
 

 

Advanced Search

Browse

All of OPARUCommunities & CollectionsPersonsInstitutionsPublication typesUlm SerialsDewey Decimal ClassesEU projects UlmDFG projects UlmOther projects Ulm

My Account

LoginRegister

Statistics

View Usage Statistics

Policy | kiz service OPARU | Contact Us
Impressum | Privacy statement