Evaluation of a novel subtilisin inhibitor gene and mutant derivatives for the expresion and secretion of mouse tumor necrosis factor alpha by Streptomyces lividans

Article Abstract:

The complementary DNA of the murine tumor necrosis factor (mTNF) was fused with the signal sequence of the gene encoding novel subtilisin inhibitor (VSI) of Streptomyces venezuelae. Immunoblot analysis of the recombinant mTNF produced by S. lividans transformants with mutated VSI signal peptides indicated the presence of varied levels of mTNF secretion that was altered by the effect of positive charges in the N regions on the signal peptides. However, the presence of positive charges in the N region was not an integral component of protein export.

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Stable Escherichia coli-Clostridium acetobutylicum shuttle vector for secretion of murine tumor necrosis factor alpha

Article Abstract:

Cloning of mRNF-alpha cDNA in a stable Escherichia coli-Clostridium acetobutylicum shuttle vector is described. Controls confirmed presence and stability of the recombinant plasmids and expression and secretion of murine tumor necrosis factor alpha (mTNF-alpha) were monitored during growth. Investigations of elaboration of a gene transfer system for cancer treatment using anaerobic bacteria are detailed.

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Insertion or deletion of the Cheo box modifies radiation inducibility of Clostridium promoters

Article Abstract:

Introducing an extra Cheo box into the promoter region of the tumor necrosis factor alpha gene in genetically engineered clostridia can increase the secretion of the protein when the bacterium is exposed to radiation. This system could be used to treat cancer patients because the treatment could be stopped at any time by killing the bacterium with antibiotics.

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