Difference between revisions of "Part:BBa K1021020"
(→Usage and Biology) |
|||
| Line 7: | Line 7: | ||
===Usage and Biology=== | ===Usage and Biology=== | ||
*The part was constructed from BBa_I712074 and BBa_E0040. | *The part was constructed from BBa_I712074 and BBa_E0040. | ||
| + | *This construct was modified to contain a bacterial RBS so that we could characterize its transcriptional functionality. A fluorescence assay was run using a 96 well plate as described in the protocol section. T7+RBS+GFP and T7+GFP (BBa_K1021020) constructs in <i>E. coli</i> BL21-A1 were examined for their relative fluorescence (fluorescence/OD600). A T7 construct in <i>E. coli</i> BL21-A1 without downstream elements was utilized as a control along with the use of the aforementioned constructs in <i>E. coli</i> DH5α. As can be seen from the graph there were evident differences in peak fluorescence between the T7+RBS+GFP construct and the T7+GFP construct and controls. The T7+GFP+RBS construct appears to have a 20 fold higher relative fluorescence than the T7+GFP construct or the controls. | ||
| + | <br> | ||
| + | [[File:Fluorreal.png]] | ||
| + | |||
<!-- --> | <!-- --> | ||
Latest revision as of 03:56, 29 October 2013
PT7+GFP
The gene for green fluorescent protein sits downstream of the T7 constitutive promoter. This gene can be naturally expressed in Escherichia coli BL21-A1 in conjunction with T7 polymerase and can be expressed in Escherichia coli DH5α after co-transformation.
Usage and Biology
- The part was constructed from BBa_I712074 and BBa_E0040.
- This construct was modified to contain a bacterial RBS so that we could characterize its transcriptional functionality. A fluorescence assay was run using a 96 well plate as described in the protocol section. T7+RBS+GFP and T7+GFP (BBa_K1021020) constructs in E. coli BL21-A1 were examined for their relative fluorescence (fluorescence/OD600). A T7 construct in E. coli BL21-A1 without downstream elements was utilized as a control along with the use of the aforementioned constructs in E. coli DH5α. As can be seen from the graph there were evident differences in peak fluorescence between the T7+RBS+GFP construct and the T7+GFP construct and controls. The T7+GFP+RBS construct appears to have a 20 fold higher relative fluorescence than the T7+GFP construct or the controls.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 696