Regulatory

Part:BBa_J100534:Design

Designed by: Cathleen Krabak, Lindsey Voelker, Kate Kilpatrick, Eboni Thomas   Group: Campbell M Lab   (2020-02-09)
Revision as of 20:32, 13 February 2020 by Cakrabak (Talk | contribs)


Constitutive RFP Promoter in E. coli


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 35
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We decided not to order the AHL chemical because there is no lux cassette in the E. coli cultures that we will use (not possible for the lux gene to be expressed). Therefore, we are allowing it to function as a constitutive promoter. We have two experimental groups that will be compared to a negative control, which contains water instead of the diluted oligo solution, and a positive control, which contains neither water nor the oligo solution. Spectrophotometry will be used to compare the optical density of each E. coli population and to create a ratio of red to optical density for the purpose of measuring the amount of red in each cell. This will tell us how effective the promoter is. We will alter several variables, including temperature, to se if they effect our results. We are currently looking into whether or not there is a gene that is similar to the lux R gene naturally expressed in E. coli that our promoter might be able to affect/function with.


Source

This promoter sequence came from an article published on NCBI: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4444344/


References