Difference between revisions of "Part:BBa K314200:Experience"

Latest revision as of 01:38, 16 October 2019

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.

User Reviews

UNIQc2e521309d244bc3-partinfo-00000000-QINU UNIQc2e521309d244bc3-partinfo-00000001-QINU

@@ Line 1: / Line 1: @@
 __NOTOC__
-This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
+===Evidence that Tse2 works as expected===
-how you used this part and how it worked out.
+'''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===
-If Tse2 is combined with a functioning Type 6 secretion system and [https://parts.igem.org/wiki/index.php?title=Part:BBa_K314201 Tsi2] it results in a killing system that can be targeted to gram negative bacteria. This could be used in a probiotic application.
+Tse2 is a potent bacterial toxin and thus has many applications, with or without its immunity protein, [https://parts.igem.org/Part:BBa_K314201 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 [https://parts.igem.org/wiki/index.php?title=Part:BBa_K314201 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].
 <br>
 <br>
-[[Image:Washington_Type_VI_secretion_image.png]]
+[[Image:Washington_Type_VI_secretion_image.png|frame|center|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.
-Tse2 could also be used as a death gene similar to ccdb.
+ [[Image:Tse2021.jpg|800px|Frame|center| Growth curve ]]
 ===User Reviews===