Difference between revisions of "Part:BBa K2447017"

Latest revision as of 08:30, 1 November 2017

Constitutive E2 killing protein production

As part of the double plasmid system to activate the kill switch, this second plasmid (if this part is integrated into a suitable backbone like pE2K) will be constitutively producing the E2 Colicin protein, and actively killing the cells. Depending on the amount of IM2 proteins produced by the first plasmid (BBa_K2447016), the cells would still survive when the kill switch is not in the 'ON' state (low phosphate (close to 0 uM) and low temperature (below 36 degrees) condition).

This construct is useful to serve as a highly specific kill switch where under high phosphate concentration and/or when the temperature is higher than 36 degrees, there would be constant production of E2 immunity protein to sequester the killing protein E2. For the remaining scenario such as when the phosphate concentration is low and when temperature is lower than 36 degrees, the kill-switch would be 'activated' where the production of E2 immunity protein will be down-regulated causing accumulation of E2 Colicin and subsequent cell death.

For more information, do visit our [http://2017.igem.org/Team:NUS_Singapore/Experiments experimental page] on wiki.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 7
Illegal NheI site found at 30
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 89
1000
COMPATIBLE WITH RFC[1000]

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 <partinfo>BBa_K2447017 short</partinfo>
-This design of this cascade system is similar to our other part [https://parts.igem.org/wiki/index.php?title=Part:BBa_K2447016 BBa_K2447016], with the only exception where the GFP coding sequence is being replaced by E2 immunity protein.
+As part of the double plasmid system to activate the kill switch, this second plasmid (if this part is integrated into a suitable backbone like pE2K) will be constitutively producing the E2 Colicin protein, and actively killing the cells. Depending on the amount of IM2 proteins produced by the first plasmid ([https://parts.igem.org/wiki/index.php?title=Part:BBa_K2447016 BBa_K2447016]), the cells would still survive when the kill switch is not in the 'ON' state (low phosphate (close to 0 uM) and low temperature (below 36 degrees) condition).
-With the addition of another plasmid that is capable of constitutive production of E2 Colicin (an DNAse), this construct is useful to serve as a highly specific kill-switch where under normal circumstances (high phosphate concentrations or when the temperature is higher than 36 degrees), there would be constant production of E2 immunity protein to sequester the killing protein E2. For other scenarios such as when the phosphate concentration is low and when temperature is lower than 36 degree, the kill-switch would be 'activated' where downstream production of E2 immunity protein will be down-regulated and causing over accumulation of E2 Colicin and subsequent cell death.
+This construct is useful to serve as a highly specific kill switch where under high phosphate concentration and/or when the temperature is higher than 36 degrees, there would be constant production of E2 immunity protein to sequester the killing protein E2. For the remaining scenario such as when the phosphate concentration is low and when temperature is lower than 36 degrees, the kill-switch would be 'activated' where the production of E2 immunity protein will be down-regulated causing accumulation of E2 Colicin and subsequent cell death.
+For more information, do visit our [http://2017.igem.org/Team:NUS_Singapore/Experiments experimental page] on wiki.
 <!-- Add more about the biology of this part here