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Title: Stable Escherichia coli-Clostridium acetobutylicum shuttle vector for secretion of murine tumor necrosis factor alpha
Authors: Theys, Jan ×
Nuyts, Sandra
Landuyt, W
Van Mellaert, Lieve
Dillen, Christiane
Böhringer, M
Dürre, P
Lambin, Philippe
Anné, Jozef #
Issue Date: Oct-1999
Series Title: Applied and Environmental Microbiology vol:65 issue:10 pages:4295-300
Abstract: Recombinant plasmids were constructed to secrete mouse tumor necrosis factor alpha (mTNF-alpha) from Clostridium acetobutylicum. The shuttle plasmids contained the clostridial endo-beta1, 4-glucanase (eglA) promoter and signal sequence that was fused in frame to the mTNF-alpha cDNA. The construction was first tested in Escherichia coli and then introduced in C. acetobutylicum DSM792 by electroporation. Controls confirmed the presence and stability of the recombinant plasmids in this organism. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and an in vitro cytotoxic assay were used to monitor expression and secretion of mTNF-alpha during growth. Significant levels of biologically active mTNF-alpha were measured in both lysates and supernatants. The present report deals with investigations on the elaboration of a gene transfer system for cancer treatment using anaerobic bacteria.
ISSN: 0099-2240
Publication status: published
KU Leuven publication type: IT
Appears in Collections:Laboratory of Molecular Bacteriology (Rega Institute)
Laboratory of Experimental Radiotherapy
× corresponding author
# (joint) last author

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