Difference between revisions of "Part:BBa K2967025"
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<partinfo>BBa_K2967025 short</partinfo> | <partinfo>BBa_K2967025 short</partinfo> | ||
| − | + | Improvement of Pyear | |
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| + | This year, we chose BBa_K (PyeaR-Luc) as an alternative to our inflammatory sensor, due to promoter PyeaR is sensitive to nitrate and nitrite. When nitrate and nitrite enter E. coli, they will be converted to nitric oxide. Then nitric oxide will bind to the repressor protein NsrR that inactivates PyeaR to inhibit transcription of downstream genes.[1] | ||
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| + | However, we noticed detectable basal expression (leakage) from the characterization of the most sensitive NO sensor (PyeaR-Luc) (Fig. 2A). To reduce sensor basal background, we integrated two different approaches. For the first approach, we inserted an extra NsrR binding sequence (NsrRBS) downstream of PyeaR to create a ‘roadblocking’ effect [2] (Fig. 1). Compare to the unmodified Pyear-luc system (Fig.2B), the histogram of luminescence data demonstrated that the relative lower luciferase signal in Pyear-NsrRBS system in the absence of NO. | ||
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===Usage and Biology=== | ===Usage and Biology=== | ||
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| − | <span class= | + | <span class="h3bb">Sequence and Features</span> |
<partinfo>BBa_K2967025 SequenceAndFeatures</partinfo> | <partinfo>BBa_K2967025 SequenceAndFeatures</partinfo> | ||
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Revision as of 07:41, 3 October 2019
The yeaR promoter added an extra NsrR Binding Sequences
Improvement of Pyear
This year, we chose BBa_K (PyeaR-Luc) as an alternative to our inflammatory sensor, due to promoter PyeaR is sensitive to nitrate and nitrite. When nitrate and nitrite enter E. coli, they will be converted to nitric oxide. Then nitric oxide will bind to the repressor protein NsrR that inactivates PyeaR to inhibit transcription of downstream genes.[1]
However, we noticed detectable basal expression (leakage) from the characterization of the most sensitive NO sensor (PyeaR-Luc) (Fig. 2A). To reduce sensor basal background, we integrated two different approaches. For the first approach, we inserted an extra NsrR binding sequence (NsrRBS) downstream of PyeaR to create a ‘roadblocking’ effect [2] (Fig. 1). Compare to the unmodified Pyear-luc system (Fig.2B), the histogram of luminescence data demonstrated that the relative lower luciferase signal in Pyear-NsrRBS system in the absence of NO.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]