Part:BBa_K314200:Experience

Evidence that Tse2 works as expected

This evidence can be found on [http://2010.igem.org/Team:Washington/Gram_Negative/Test this page] from the 2010 UW iGEM Wiki, under the heading "Evidence that Tse2 and Tsi2 are Function As Expected"

Applications of BBa_K314200

Tse2 is a potent bacterial toxin and thus has many applications, with or without its immunity protein, Tsi2. One potential application is use as a biotechnology agent in the way that the CcdB, the Death Protein, is used.

Tse2 could also be combined with the T6SS to result in a system that targets this toxin into the cytoplasm of target Gram-negative bacteria. This is useful to create Gram-negative bacteria that have been directed to target other Gram-negative bacteria for killing in response to a user-defined signal.

If Tse2 is combined with a functioning Type 6 Secretion System and the antitoxin Tsi2 it results in a killing system that can be targeted to Gram-negative bacteria. This could be used in a probiotic application. This was the goal behind the Gram-negative portion of the [http://2010.igem.org/Team:Washington 2010 UW iGEM project].

Diagram of the Tse2/Tsi2/ Type VI Secretion System as it would be used in a probiotic application

The Trieste iGEM team 2012 explored the possibility to use this toxin for the horizontal gene transfer regulation system (gene guard).

First of all we needed to deeply characterize this part. we build this construct K875002(T5Lac Op) - B0031(RBS) - K314200(Tse2 toxin) - B0015(double terminator). A growth assay has been done. Here DH5alpha E.coli bacteria transformed with the toxin under the IPTG inducible promoter were grown for several hour in LB media. Six samples with a OD 600 absorbance of 0.1 were divided into two group: untreated and treated with IPTG 1mM at time zero.

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