Team:Washington/Gram Negative/Design

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Revision as of 01:07, 9 September 2010

In order to create a probiotic application for this system, we first attempt to express it heterologously in E. Coli.  Starting from a Fosmid containing our T6SS, we are using Recombineering as described by Warming et al (2005) to replace the strict native regulation with robust T7 promoters to create strong expression of the T6SS.

At the same time, we are creating inducible circuits to allow for production and regulation for the antibacterial toxin and antitoxin system, allowing us to create "assassin cells" upon induction.

← Overview of the Gram(-) Therapeutic Building the Gram(-) Therapeutic →

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-The ''Pseudomonas aeruginosa'' genome includes a Type VI Secretion System (T6SS) which has been shown to target prokaryotic cells with a toxin (Hood, 2010).  In order to create a probiotic application for this system, we first attempt to express it heterologously in ''E. Coli''.
+  In order to create a probiotic application for this system, we first attempt to express it heterologously in ''E. Coli''.  Starting from a Fosmid containing our T6SS, we are using Recombineering as described by Warming et al (2005) to replace the strict native regulation with robust T7 promoters to create strong expression of the T6SS.
-Using Recombineering as described by Warming et al (2005), we are replacing the strict native regulation with robust T7 promoters to create strong expression of the T6SS.
 At the same time, we are creating inducible circuits to allow for production and regulation for the antibacterial toxin and antitoxin system, allowing us to create "assassin cells" upon induction.