Part:BBa_K1486048
CpxR reporter
Calgary's BBa_K339007 [1] repaired: the part didn't have the sequence they said they had, it was missing most of the promoter.
Biology of the Part
E coli have several native sigma factors in the genome that are upregulated during envelope stress. There is also a new regulon that is currently studied widely, called the Cpx regulon, a well characterized periplasmic regulon found in E. coli. The Cpx pathway is involved in maintaining cellular adhesion and keeping periplasmic environment in check. It regulates proteins which are responsible for cellular adhesion and pili formation. The Cpx pathway is activated by factors which cause protein aggregation in the periplasmic space of gram negative bacteria. Extracellular stresses such as exposure to ethanol, change in pH and temperature change induce the activation of the Cpx heat shock regulon. In our BioBrick this should increase the transcription of the RFP which it regulates.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 636
Illegal AgeI site found at 748 - 1000COMPATIBLE WITH RFC[1000]
Characterisation
To check if the cells where responding to different kinds of stress we put them in different environments: 40 mM KCl, 80 mM KCl, 5 % ethanol and 10 % ethanol . As a negative control we added PBS to the same cells. As an other negative control cells that did not have the RFP coding sequence where put in the same environments.
http://2014.igem.org/File:BBa_K3396007_copy.png
//classic/regulatory/uncategorized
//direction/forward
//function/reporter/fluorescence
//promoter
//regulation/positive
regulator
transcriptional
biology | |
chassis | Escherichia coli |
color | Red |
device_type | reporter |
direction | Forward |
emission | 632 |
excitation | 555 |
resistance | Chloramphenicol (backbone) |