Team:Paris Bettencourt/Experiments/Methodologies/K090403J

From 2011.igem.org

(Difference between revisions)
 
(12 intermediate revisions not shown)
Line 1: Line 1:
{{:Team:Paris_Bettencourt/tpl_test}}
{{:Team:Paris_Bettencourt/tpl_test}}
 +
<h1>Integration plasmid</h1>
-
== Cyrille ==
+
In parallel to the transformation with the replicative plasmid (pHM3), we tried to use a Bacillus subtilis integration plasmid. For that, we used the construction of the B.subtilis integration vector pSBINT1 built by the 2008 Cambridge iGEM team.
-
 
+
-
Control digestion of pHM3 in NcoI
+
-
 
+
-
[[File:CP2009.jpg|thumb|center]]
+
-
 
+
-
Control of the size of the insert
+
-
 
+
-
[[File:CP2009_XP.jpg|thumb|center|S75 *5; S82 *5; 104 *5]]
+
-
[[File:CP2009_XP_ComS.jpg|thumb|center|ComS *3]]
+
-
 
+
-
== Baptiste, Hovannes & Ouriel ==
+
-
 
+
-
===Characterisation of T7 autoloop, RFP - GFP ===
+
-
 
+
-
We characterized T7 autoloop by microscopy.  
+
-
Leak,
+
-
 
+
-
 
+
{| border="1" class="wikitable" style="text-align: center;"
{| border="1" class="wikitable" style="text-align: center;"
-
|+T7 autoloop RFP+ Control / E-coli at 37°C
+
|+ Bacillus subtilis integration vector
|-
|-
-
|[[File:T7 autoloop ctrl-7.jpg|290px|thumb|center|E-Coli  at 37°C  (trans image)]]
+
|[[File:PBSINT1_final.png|900px|thumb|center|]]
-
|[[File:T7 autoloop ctrl-8.jpg|290px|thumb|center|E-Coli  at 37°C  (gfp image)]]
+
-
|[[File:T7 autoloop ctrl-9.jpg|290px|thumb|center|E-Coli  at 37°C  (rfp image)]]
+
-
|-
+
-
|[[File:T7 autoloop ctrl-10.jpg|290px|thumb|center|E-Coli  at 37°C  (trans image)]]
+
-
|[[File:T7 autoloop ctrl-11.jpg|290px|thumb|center|E-Coli  at 37°C  (gfp image)]]
+
-
|[[File:T7 autoloop ctrl-12.jpg|290px|thumb|center|-Coli  at 37°C  (rfp image)]]
+
-
|}
+
-
 
-
TEXT
 
-
 
-
 
-
{| border="1" class="wikitable" style="text-align: center;"
 
-
|+T7 autoloop full no terminator / E-coli at 37°C
 
|-
|-
-
|[[File:T7 autoloop ctrl-25.jpg|290px|thumb|center|E-Coli  at 37°C  (trans image)]]
 
-
|[[File:T7 autoloop ctrl-26.jpg|290px|thumb|center|E-Coli  at 37°C  (gfp image)]]
 
-
|[[File:T7 autoloop ctrl-27.jpg|290px|thumb|center|E-Coli  at 37°C  (rfp image)]]
 
-
|-
 
-
|[[File:T7 autoloop ctrl-31.jpg|290px|thumb|center|E-Coli  at 37°C  (trans image)]]
 
-
|[[File:T7 autoloop ctrl-32.jpg|290px|thumb|center|E-Coli  at 37°C  (gfp image)]]
 
-
|[[File:T7 autoloop ctrl-33.jpg|290px|thumb|center|-Coli  at 37°C  (rfp image)]]
 
|}
|}
-
 
+
<p>At first, we managed to clone each of our parts in the 5' integration sequence for the AmyE locus of B.subtilis (<partinfo>K143001</partinfo>).</p>
-
{| border="1" class="wikitable" style="text-align: center;"
+
<p>After, we want to clone them into the integrative plasmid (<partinfo>K090403</partinfo>).</p>
-
|+T7 autoloop full with  terminator / E-coli at 37°C
+
-
|-
+
-
|[[File:T7_loop_1.jpg|290px|thumb|center|E-Coli  at 37°C  (trans image)]]
+
-
|[[File:T7 loop 1 gfp.jpg|290px|thumb|center|E-Coli  at 37°C  (gfp image)]]
+
-
|[[File:T7 loop 1 rfp.jpg|290px|thumb|center|E-Coli  at 37°C  (rfp image)]]
+
-
|-
+
-
|[[File:T7 loop 2.jpg|290px|thumb|center|E-Coli  at 37°C  (trans image)]]
+
-
|[[File:T7 loop 1 gfp.jpg|290px|thumb|center|E-Coli  at 37°C  (gfp image)]]
+
-
|[[File:T7 loop 2 rfp.jpg|290px|thumb|center|-Coli  at 37°C  (rfp image)]]
+
-
|}
+
-
 
+
-
===YFP-tetR===
+
-
 
+
-
We
+
-
 
+
-
 
+
-
TEXT
+
<html>
<html>

Latest revision as of 02:34, 22 September 2011

Team IGEM Paris 2011

Integration plasmid

In parallel to the transformation with the replicative plasmid (pHM3), we tried to use a Bacillus subtilis integration plasmid. For that, we used the construction of the B.subtilis integration vector pSBINT1 built by the 2008 Cambridge iGEM team.

Bacillus subtilis integration vector
PBSINT1 final.png

At first, we managed to clone each of our parts in the 5' integration sequence for the AmyE locus of B.subtilis (<partinfo>K143001</partinfo>).

After, we want to clone them into the integrative plasmid (<partinfo>K090403</partinfo>).

Retrieved from "http://2011.igem.org/Team:Paris_Bettencourt/Experiments/Methodologies/K090403J"