Difference between revisions of "Part:BBa K381001"
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Nitrate and Nitrite sensitive promoter PyeaR with a GFP coding device and strong RBS to create a nitrate-sensitive system which signals through expression of GFP. | Nitrate and Nitrite sensitive promoter PyeaR with a GFP coding device and strong RBS to create a nitrate-sensitive system which signals through expression of GFP. | ||
− | Optimum use appears to be when sensing nitrate within a range of 0 - 20 mM | + | PyeaR is normally repressed by NsrR, a protein native to most 'E. coli' cells. When Nitrate or Nitrite enter the cell it is converted to Nitric Oxide. This binds to NsrR, halting the repression and allowing the production of GFP. |
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+ | Optimum use appears to be when sensing nitrate within a range of 0 - 20 mM (Calculated from Miller assays) | ||
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+ | Activity between 0 - 10mM was investigated using an assay measuring change in GFP expression with increasing levels of Potassium Nitrate, the results of which can be seen below. | ||
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+ | [[Image:GFPChar0to10.png|centre|500px]] | ||
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<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here |
Revision as of 11:35, 26 October 2010
Nitrate reporter: PyeaR - GFP composite
Nitrate and Nitrite sensitive promoter PyeaR with a GFP coding device and strong RBS to create a nitrate-sensitive system which signals through expression of GFP.
PyeaR is normally repressed by NsrR, a protein native to most 'E. coli' cells. When Nitrate or Nitrite enter the cell it is converted to Nitric Oxide. This binds to NsrR, halting the repression and allowing the production of GFP.
Optimum use appears to be when sensing nitrate within a range of 0 - 20 mM (Calculated from Miller assays)
Activity between 0 - 10mM was investigated using an assay measuring change in GFP expression with increasing levels of Potassium Nitrate, the results of which can be seen below.
Sequence and Features
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Unknown
- 12INCOMPATIBLE WITH RFC[12]Unknown
- 21INCOMPATIBLE WITH RFC[21]Unknown
- 23INCOMPATIBLE WITH RFC[23]Unknown
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 773