Difference between revisions of "Part:BBa K2447012"
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TlpA36 is a temperature sensitive protein which is capable of binding to promoter pTlpA. Under temperature of above 36 degrees Celsius, TlpA36 protein would stay in monomeric form and hence it loses the ability to bind to the promoter pTlpA36. If the temperature drops below 36 degree Celsius, TlpA36 proteins would form dimers; which are capable of repressing the pTlpA promoter and prevent the recruitment of RNA polymerase to repress downstream gene expression. | TlpA36 is a temperature sensitive protein which is capable of binding to promoter pTlpA. Under temperature of above 36 degrees Celsius, TlpA36 protein would stay in monomeric form and hence it loses the ability to bind to the promoter pTlpA36. If the temperature drops below 36 degree Celsius, TlpA36 proteins would form dimers; which are capable of repressing the pTlpA promoter and prevent the recruitment of RNA polymerase to repress downstream gene expression. | ||
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| + | RDFZ-China 2018 Experience | ||
| + | While we are designing our drug delivery bacteria project, we found out that TlpA has too high activation energy and TlpA36 is too low comparing to human body temperature. So, we found this TlpA39 on Shapiro’s lab, and utilized it into our project. | ||
| + | We also constructed a TlpA39-LVA to see if the LVA tag will accelerate the derepression process, but we found that there is no repression to TlpA39 since it has been rapidly degraded. | ||
| + | First we characterized a set of thermal repressors including K2447012 and our new improved part K2572000. | ||
| + | https://static.igem.org/mediawiki/2018/thumb/8/8e/T--RDFZ-China--TSRquali.jpeg/800px-T--RDFZ-China--TSRquali.jpeg | ||
| + | We can see that at 35 degree Celsius, TlpA36-K2447012 starts to derepress. At 37 degree Celsius, TlpA39-K2572000 starts to derepress. At 39.5 degree Celsius, Tcl42-K2572001 starts to express. | ||
| + | Then we characterized it on plate reader under different temperature. | ||
| + | |||
| + | https://static.igem.org/mediawiki/2018/6/66/T--RDFZ-China--TSR.png | ||
| + | Which clearly shows the difference in expression under different temperature. | ||
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Latest revision as of 23:22, 17 October 2018
Coding region for temperature sensitive protein TlpA36
TlpA36 is a temperature sensitive protein which is capable of binding to promoter pTlpA. Under temperature of above 36 degrees Celsius, TlpA36 protein would stay in monomeric form and hence it loses the ability to bind to the promoter pTlpA36. If the temperature drops below 36 degree Celsius, TlpA36 proteins would form dimers; which are capable of repressing the pTlpA promoter and prevent the recruitment of RNA polymerase to repress downstream gene expression.
RDFZ-China 2018 Experience
While we are designing our drug delivery bacteria project, we found out that TlpA has too high activation energy and TlpA36 is too low comparing to human body temperature. So, we found this TlpA39 on Shapiro’s lab, and utilized it into our project.
We also constructed a TlpA39-LVA to see if the LVA tag will accelerate the derepression process, but we found that there is no repression to TlpA39 since it has been rapidly degraded.
First we characterized a set of thermal repressors including K2447012 and our new improved part K2572000.
We can see that at 35 degree Celsius, TlpA36-K2447012 starts to derepress. At 37 degree Celsius, TlpA39-K2572000 starts to derepress. At 39.5 degree Celsius, Tcl42-K2572001 starts to express.
Then we characterized it on plate reader under different temperature.
Which clearly shows the difference in expression under different temperature.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 508
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 793
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1096
- 1000COMPATIBLE WITH RFC[1000]