Difference between revisions of "Part:BBa K2275011"

 
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<partinfo>BBa_K2275011 short</partinfo>
 
<partinfo>BBa_K2275011 short</partinfo>
  
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in front of RBS
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A promoter PyeaR can be induced when it senses the nitrate and nitrite (Lin, et al., 2007). The BBCS-Bristol 2010 iGEM team constructed the biobrick K381001, which is composed of a PyeaR promoter, a strong RBS B0030 and a reporter gene GFP. It can emit green fluorescence when detecting nitrate so that it can also be served as a nitrate sensor.
 +
PyeaR is repressed by NsrR protein under no nitrate or nitric oxide condition, and is activated when nitrate or nitrite is existing. In theory, the biobrick K381001 can’t emit fluorescence under no nitrate or nitrite. However, the data showed that it can still be activated. In order to improve the biobrick K381001, we decided to add an additional nsrR sequence to it so as to reinforce repression and decrease interference. As a result, fluorescence basal level can be decreased, and detection will be enhanced.
 +
 
 +
Functional test:
 +
By adding an nsrR sequence to K381001 before RBS, we successfully decreased the fluorescence basal level. That is to say, under no nitrate induction, we can get lower fluorescence and maintain ability to do more accurate and easier nitrate detection without interference by other factors.
 +
[[File:Functional test of PyeaR-NsrR binding site-B0030.png|350px|centre]]
 +
[[File:Sensitivity to nitrate after 12 hours induction.png|350px|centre]]
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 16:34, 27 October 2017


PyeaR-NsrR binding site-B0030-GFP composite

A promoter PyeaR can be induced when it senses the nitrate and nitrite (Lin, et al., 2007). The BBCS-Bristol 2010 iGEM team constructed the biobrick K381001, which is composed of a PyeaR promoter, a strong RBS B0030 and a reporter gene GFP. It can emit green fluorescence when detecting nitrate so that it can also be served as a nitrate sensor. PyeaR is repressed by NsrR protein under no nitrate or nitric oxide condition, and is activated when nitrate or nitrite is existing. In theory, the biobrick K381001 can’t emit fluorescence under no nitrate or nitrite. However, the data showed that it can still be activated. In order to improve the biobrick K381001, we decided to add an additional nsrR sequence to it so as to reinforce repression and decrease interference. As a result, fluorescence basal level can be decreased, and detection will be enhanced.

Functional test: By adding an nsrR sequence to K381001 before RBS, we successfully decreased the fluorescence basal level. That is to say, under no nitrate induction, we can get lower fluorescence and maintain ability to do more accurate and easier nitrate detection without interference by other factors.

Functional test of PyeaR-NsrR binding site-B0030.png
Sensitivity to nitrate after 12 hours induction.png

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 107
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 802