Difference between revisions of "Part:BBa K817013"
(2 intermediate revisions by the same user not shown) | |||
Line 2: | Line 2: | ||
<partinfo>BBa_K817013 short</partinfo> | <partinfo>BBa_K817013 short</partinfo> | ||
− | As we sought to design a thermal sensitive composite part, we integrate our modified pCI promoter(modification of pCI, BBa_R0051) with temperature sensitive CI repressor, which is cloned from Harvard’s thermal regulation part, 2008. (BBa_K098988). We showed the modified promoter design had superior repression effects. | + | As we sought to design a thermal sensitive composite part, we integrate our modified pCI promoter(modification of pCI, [[Part:BBa_R0051|BBa_R0051]]) with temperature sensitive CI repressor, which is cloned from Harvard’s thermal regulation part, 2008. ([[Part:BBa_K098988|BBa_K098988]]). We showed the modified promoter design had superior repression effects. |
+ | |||
+ | ==Modified P<sub>CI</sub>== | ||
+ | <p style="text-indent: 2em;">The circuit incorporates a temperature sensitive cI promoter(CIts) to sense the temperature upshift. In our test we use a thermal adjustable plate reader to detect the mRFP flurorescence. In the beginning, we keep the temperature under 30 Celsius degrees for over 1 hour, and then detect the emission of mRFP. As the dimerized CIts repressor in lower temperature would specifically bind and repress P<sub>CI</sub>, and further hinder the expression of mRFP. We can expect the emission to be low under 610 nm wavelength. We then abruptly increase our temperature to 37 celsius degrees, as we expect the CIts dimer would decompose and lose the function of repressing mRFP expression. </p> | ||
+ | |||
+ | <center><html><img src='https://static.igem.org/mediawiki/2012/thumb/c/cb/NTU-Taida-Result-Thermal-pCI.png/800px-NTU-Taida-Result-Thermal-pCI.png' width='600px'></html></center> | ||
+ | |||
+ | <p style="text-indent: 2em;">The result showed a low level of mRFP expression under wavelength of 580 nm (excitation) and 610 nm (emission). After the sudden temperature upshift, the expression of mRFP steadily rises, and results in 5.7 folds increase in the 8th hour after the temperature upshift. </p> | ||
+ | <p style="text-indent: 2em;">This proved the fact that repressor CIts and P<sub>CI</sub> can largely lead to increase in protein expression, and can be used in our circuit design as it may turn on the circuit inside human body and spontaneously close down after the bacteria is expelled out. </p> | ||
+ | |||
+ | We put our experience back to the related pages on wiki: | ||
+ | |||
+ | # Modified part: [[https://parts.igem.org/Part:BBa_K098988:Experience BBa_K098988:Experience]] | ||
+ | # Modified basic part: [[https://parts.igem.org/Part:BBa_R0051:Experience BBa_R0051:Experience]] | ||
− | |||
− | |||
<!-- --> | <!-- --> |
Latest revision as of 02:08, 6 October 2012
pCI(OR1-OL1)-mRFP
As we sought to design a thermal sensitive composite part, we integrate our modified pCI promoter(modification of pCI, BBa_R0051) with temperature sensitive CI repressor, which is cloned from Harvard’s thermal regulation part, 2008. (BBa_K098988). We showed the modified promoter design had superior repression effects.
Modified PCI
The circuit incorporates a temperature sensitive cI promoter(CIts) to sense the temperature upshift. In our test we use a thermal adjustable plate reader to detect the mRFP flurorescence. In the beginning, we keep the temperature under 30 Celsius degrees for over 1 hour, and then detect the emission of mRFP. As the dimerized CIts repressor in lower temperature would specifically bind and repress PCI, and further hinder the expression of mRFP. We can expect the emission to be low under 610 nm wavelength. We then abruptly increase our temperature to 37 celsius degrees, as we expect the CIts dimer would decompose and lose the function of repressing mRFP expression.
![](https://static.igem.org/mediawiki/2012/thumb/c/cb/NTU-Taida-Result-Thermal-pCI.png/800px-NTU-Taida-Result-Thermal-pCI.png)
The result showed a low level of mRFP expression under wavelength of 580 nm (excitation) and 610 nm (emission). After the sudden temperature upshift, the expression of mRFP steadily rises, and results in 5.7 folds increase in the 8th hour after the temperature upshift.
This proved the fact that repressor CIts and PCI can largely lead to increase in protein expression, and can be used in our circuit design as it may turn on the circuit inside human body and spontaneously close down after the bacteria is expelled out.
We put our experience back to the related pages on wiki:
- Modified part: [BBa_K098988:Experience]
- Modified basic part: [BBa_R0051:Experience]
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Unknown
- 23INCOMPATIBLE WITH RFC[23]Unknown
- 25INCOMPATIBLE WITH RFC[25]Unknown
- 1000COMPATIBLE WITH RFC[1000]