Difference between revisions of "Part:BBa M50470"

Revision as of 14:43, 12 December 2018

Cold Shock Sensor w/ aeBlue+3X-FLAG tag Pigment Reporter

This plasmid senses cold temperatures using the cold-shock protein A promoter (cspA) and produces blue pigment (aeBlue) when activated. Ideally, this system should work such that the colder the temperature, the more blue pigment that is produced. aeBlue is tagged with a 3X-FLAG tag, which facilitates Western blot analysis of aeBlue production. Dynamic range experimentation concluded that no meffRed is visibly produced after cold-shock at a variety of warm temperatures after 60 minutes; Western blot analysis as well as plate observations reveal that aeBlue is in fact produced, however it takes an extended amount of time and/or does not produce enough to be visibly seen. Future recommendations include designing a better way to facilitate cold shock response for our intended project.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

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 <partinfo>BBa_M50470 short</partinfo>
-This is a cold-shock temperature sensor that reflects low temperatures through aeBlue chromoprotein expression. At low temperatures, roughly 15C and below, the cspA promoter allows transcription of the chromoprotein, and at high temperatures the promoter is not active, so no blue  The level of GFP expression corresponds to the level of atrazine in the surrounding environment.
+This plasmid senses cold temperatures using the cold-shock protein A promoter (cspA) and produces blue pigment (aeBlue) when activated. Ideally, this system should work such that the colder the temperature, the more blue pigment that is produced. aeBlue is tagged with a 3X-FLAG tag, which facilitates Western blot analysis of aeBlue production.
+Dynamic range experimentation concluded that no meffRed is visibly produced after cold-shock at a variety of warm temperatures after 60 minutes; Western blot analysis as well as plate observations reveal that aeBlue is in fact produced, however it takes an extended amount of time and/or does not produce enough to be visibly seen. Future recommendations include designing a better way to facilitate cold shock response for our intended project.
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