Difference between revisions of "Part:BBa K2918032"
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A level 1 with 0.5 T7 promoter, a universal RBS, a GFP and a T7 terminator | A level 1 with 0.5 T7 promoter, a universal RBS, a GFP and a T7 terminator | ||
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<partinfo>BBa_K2918032 SequenceAndFeatures</partinfo> | <partinfo>BBa_K2918032 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. | ||
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| + | <div><ul> | ||
| + | <li style="display: inline-block;"> [[File:T--TUDelft--0.5gfp-nomM.png|thumb|none|444px|<b>Figure 3A:</b> The growth curve of GFP under a 0.5 T7 promoter in hours with 1 mM IPTG induction]] </li> | ||
| + | <li style="display: inline-block;"> [[File:T--TUDelft--0.5gfp-1mM.png|thumb|none|444px|<b>Figure 3B:</b> The growth curve of GFP under a 0.5 T7 promoter in hours with 1 mM IPTG induction]] </li> | ||
| + | <li> [[File:T--TUDelft--0.5gfp-10mM.png|thumb|center|444px|<b>Figure 3C:</b> The growth curve of GFP under a 0.5 T7 promoter in hours with 10 mM IPTG inductionn]] </li> | ||
| + | </ul></div> | ||
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| + | <!-- Add more about the biology of this part here | ||
| + | ===Usage and Biology=== | ||
<!-- Uncomment this to enable Functional Parameter display | <!-- Uncomment this to enable Functional Parameter display | ||
Revision as of 08:53, 4 October 2019
Medium T7 promoter - Universal RBS - GFP - T7 terminator
A level 1 with 0.5 T7 promoter, a universal RBS, a GFP and a T7 terminator
Sequence and Features
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 465
- 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 465
- 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 465
Illegal BamHI site found at 674 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 465
- 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 465
- 1000COMPATIBLE 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.
