Difference between revisions of "Part:BBa K3962341"

Latest revision as of 07:48, 16 October 2021

arabinose-mediated inducible expression of toxin Hok

This construct is composed of L-arabinose-mediate inducible promoter pBAD and toxin Hok. Hok/sok is a type I toxin/antitoxin system originating from E. coli. In our project, we used this construct to test the toxicity of Hok, which provided us with more insights to regulate the toxin-antitoxin system for preventing horizontal gene transfer.

2021 Team Leiden

https://2021.igem.org/Team:Leiden

In our project, we used this construct to test the toxicity of the Hok toxin, which provided us with more insights in regulating the balance of the toxin-antitoxin system for preventing horizontal gene transfer. The toxicity of this kill switch has been assessed via a plate reader assay (Fig 1.).

Figure 1. Plate reader assay of E. coli TOP 10 cellular growth over time with a gradient of arabinose concentrations.

Cell growth is strongly inhibited following the induction of CcdB. The results show that CcdB is not necessarily enough to kill all cells immediately when grown in liquid LB medium. Only after the exponential growth phase, the cells start to die. High concentrations of arabinose are needed for this killing effect, but that might change in different medium conditions.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 1205
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 1144
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 979
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI site found at 961

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K3962341 short</partinfo>
-This construct is composed of L-arabinose-mediate inducible promoter pBAD and toxin Hok. Hok/sok is a type I toxin/antitoxin system originated from E. coli. In our project, we used this construct to test the toxicity of toxin Hok, which provided us with more insights to regulate the toxin-antitoxin system for preventing horizontal gene transfer.
+This construct is composed of L-arabinose-mediate inducible promoter pBAD and toxin Hok. Hok/sok is a type I toxin/antitoxin system originating from E. coli. In our project, we used this construct to test the toxicity of Hok, which provided us with more insights to regulate the toxin-antitoxin system for preventing horizontal gene transfer.
 <!-- Add more about the biology of this part here
@@ Line 14: / Line 14: @@
 https://2021.igem.org/Team:Leiden
-In our project, we used this construct to test the toxicity of the Hok toxin, which provided us with more insights in regulating the balance of the toxin-antitoxin system for preventing horizontal gene transfer. The lethality of this kill switch has been assessed via a plate reader assay (Fig 1.).
+In our project, we used this construct to test the toxicity of the Hok toxin, which provided us with more insights in regulating the balance of the toxin-antitoxin system for preventing horizontal gene transfer. The toxicity of this kill switch has been assessed via a plate reader assay (Fig 1.).
 [[Image:T-Leiden-parts-Figuer3.png|500px]]