Team:Paris Bettencourt/SID

From 2012.igem.org

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(Overview)
(Overview)
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Provide an overview of this sub-project in a couple of phrases
Provide an overview of this sub-project in a couple of phrases
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Bacteria developed mechanisms to kill other bacteria, thus reducing competition between them in the environment. Some strains of E. coli produce lethal proteins called colicins which kill other bacteria, including E. coli. Colicins are built out of three main domains which are also a point of difference among many types of colicins. First domain is responsible for binding to a receptor on a bacterial membrane, second one is responsible for translocating the protein from outside to inside of a bacteria and the third one is responsible for killing bacteria. We are interested in two types of colicins, colicin E2 and colicin D. Both of them use different binding, translocation and killing mechanisms. Colicin E2 is a DNAse and colicin D is an RNAse.
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Bacteria developed mechanisms to kill other bacteria, thus reducing competition between them  
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We hypothesize that it is possible to use binding and translocation part of colicin as a "synthetic import domain" onto which we can fuse other proteins in order to import them into bacteria. Main aim of this part of the project is to fuse the RNAse domain from colicin D to the "synthetic import domain" of colicin E2. By doing this we will obtain a toxin that targets the same receptor and translocation mechanism but kills bacteria on the level of protein synthesis by cleaving tRNA. [link to suicide group].
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in the environment. Some strains of E. coli produce lethal proteins called colicins which
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In addition to this, we fused restriction enzymes FseI and I-SceI both to the colicin E2 and colicin D "synthetic import domain". These two enzymes are used in different parts of our project [link to restriction group]. We also phused a couple of other proteins to easily test the plausibility of our "synthetic import domain" system.
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kill other bacteria, including E. coli. Colicins are built out of three main domains which
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+
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are also a point of difference among many types of colicins. First domain is responsable for
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binding to a receptor on a bacterial membrane, second one is responsable for translocating
+
-
 
+
-
the protein from outside to inside of a bacteria and the third one is responsable for
+
-
 
+
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killing bacteria. We are interested in two types of colicins, colicin E2 and colicin D. Both
+
-
 
+
-
of them use different binding, translocation and killing mechanisms. Colicin E2 is a DNAse
+
-
 
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and colicin D is a RNAse.
+
-
 
+
-
We hypothesize that it is possible to use binding and translocation part of colicin as a
+
-
 
+
-
"synthetic import domain" onto which we can phuse other proteins in order to import them
+
-
 
+
-
into bacteria. Main aim of this part of the project is to phuse the RNAse domain from colicin D to the "synthetic import domain" of colicin E2. By doing this we will obtain a toxin that targets the same receptor and translocation mechanism but kills bacteria on the level of protein synthesis by cleaving tRNA. [link to suicide group].
+
-
 
+
-
In addition to this, we phused restriction enzymes FseI and I-SceI both to the colicin E2 and colicin D "synthetic import domain". These two enzymes are used in different parts of our project [link to restriction group]. We also phused a couple of other proteins to easly test the plausibility of our "synthetic import domain" system.
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==Objectives==
==Objectives==

Revision as of 16:02, 24 September 2012


iGEM Paris Bettencourt 2012

Title

Contents

Overview

Provide an overview of this sub-project in a couple of phrases

Bacteria developed mechanisms to kill other bacteria, thus reducing competition between them in the environment. Some strains of E. coli produce lethal proteins called colicins which kill other bacteria, including E. coli. Colicins are built out of three main domains which are also a point of difference among many types of colicins. First domain is responsible for binding to a receptor on a bacterial membrane, second one is responsible for translocating the protein from outside to inside of a bacteria and the third one is responsible for killing bacteria. We are interested in two types of colicins, colicin E2 and colicin D. Both of them use different binding, translocation and killing mechanisms. Colicin E2 is a DNAse and colicin D is an RNAse.

We hypothesize that it is possible to use binding and translocation part of colicin as a "synthetic import domain" onto which we can fuse other proteins in order to import them into bacteria. Main aim of this part of the project is to fuse the RNAse domain from colicin D to the "synthetic import domain" of colicin E2. By doing this we will obtain a toxin that targets the same receptor and translocation mechanism but kills bacteria on the level of protein synthesis by cleaving tRNA. [link to suicide group].

In addition to this, we fused restriction enzymes FseI and I-SceI both to the colicin E2 and colicin D "synthetic import domain". These two enzymes are used in different parts of our project [link to restriction group]. We also phused a couple of other proteins to easily test the plausibility of our "synthetic import domain" system.

Objectives

Write here the objectives of your project


Design

Present the design of your system, both in a written form, and a schematic one.

Bettencourt Sid1.png
Bettencourt Sid2.png
Bettencourt Sid3.png

Experiments and results

Characterisation of X

Experimental setup

Describe the experiment

Results

Present your results

Testing of the system

Experimental setup

Describe the experiment

Results

Present your results

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