Difference between revisions of "Part:BBa K2918030"

Revision as of 09:11, 4 October 2019

WT T7 promoter - Universal RBS - GFP - T7 terminator

A level 1 assembly of Wild type T7 promoter, a universal RBS, GFP from the registry and T7 terminator.

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal EcoRI site found at 465
12
INCOMPATIBLE WITH RFC[12]
Illegal EcoRI site found at 465
21
INCOMPATIBLE WITH RFC[21]
Illegal EcoRI site found at 465
Illegal BamHI site found at 674
23
INCOMPATIBLE WITH RFC[23]
Illegal EcoRI site found at 465
25
INCOMPATIBLE WITH RFC[25]
Illegal EcoRI site found at 465
1000
COMPATIBLE WITH RFC[1000]

Toxicity

Our Sci-Phi 29 tool is based on four components of the Φ29 bacteriophage: DNAP, TP, p5 and p6. However, overexpression of these proteins are toxic for the cell. In order to determine the optimal expression levels of the proteins in live cells, we carried out viability assays in E. coli BL21(DE3) pLysS. The results are shown in the graphs below.

Figure 3A: The growth curve of phi 29 GFP under a WT T7 promoter in hours with no IPTG induction
Figure 3B: The growth curve of phi 29 GFP under a WT T7 promoter in hours with 0.001 mM IPTG induction
Figure 3C: The growth curve of phi 29 GFP under a WT T7 promoter in hours with 0.01 mM IPTG induction
Figure 3D: The growth curve of phi 29 GFP under a WT T7 promoter in hours with WT mM IPTG induction
Figure 3E: The growth curve of phi 29 GFP under a WT T7 promoter in hours with 1 mM IPTG induction
Figure 3F: The growth curve of phi 29 GFP under a WT T7 promoter in hours with 10 mM IPTG induction

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K2918030 short</partinfo>
-A level 1 assembly of Wild type T7 promoter, a universal RBS, GFP from the registry and T7 terminator
+A level 1 assembly of Wild type T7 promoter, a universal RBS, GFP from the registry and T7 terminator.
-<!-- Add more about the biology of this part here
-===Usage and Biology===
 <!-- -->
@@ Line 12: / Line 9: @@
 <partinfo>BBa_K2918030 SequenceAndFeatures</partinfo>
+===Toxicity===
+Our Sci-Phi 29 tool is based on four components of the Φ29 bacteriophage: DNAP, TP, p5 and p6. However, overexpression of these proteins are toxic for the cell. In order to determine the optimal expression levels of the proteins in live cells, we carried out viability assays in <i>E. coli</i> BL21(DE3) pLysS. The results are shown in the graphs below.
+<div><ul>
+    <li style="display: inline-block;"> [[File:T--TUDelft--WTGFP-nomM.png|thumb|none|444px|<b>Figure 3A:</b> The growth curve of phi 29 GFP under a WT T7 promoter in hours with no IPTG induction]] </li>
+    <li style="display: inline-block;"> [[File:T--TUDelft--WTGFP-0.001mM.png|thumb|none|444px|<b>Figure 3B:</b> The growth curve of phi 29 GFP under a WT T7 promoter in hours with 0.001 mM IPTG induction]] </li>
+    <li style="display: inline-block;"> [[File:T--TUDelft--WTGFP-0.01mM.png|thumb|none|444px|<b>Figure 3C:</b> The growth curve of phi 29 GFP under a WT T7 promoter in hours with 0.01 mM IPTG induction]] </li>
+    <li style="display: inline-block;"> [[File:T--TUDelft--WTgfp-0.1mM.png|thumb|none|444px|<b>Figure 3D:</b> The growth curve of phi 29 GFP under a WT T7 promoter in hours with WT mM IPTG induction]] </li>
+    <li style="display: inline-block;"> [[File:T--TUDelft--WTGFP-1mM.png|thumb|none|444px|<b>Figure 3E:</b> The growth curve of phi 29 GFP under a WT T7 promoter in hours with 1 mM IPTG induction]] </li>
+    <li style="display: inline-block;"> [[File:T--TUDelft--WTGFP-10mM.png|thumb|none|444px|<b>Figure 3F:</b> The growth curve of phi 29 GFP under a WT T7 promoter in hours with 10 mM IPTG induction]] </li>
+    </ul></div>
+<!-- Add more about the biology of this part here
+===Usage and Biology===
 <!-- Uncomment this to enable Functional Parameter display