Reporter

Part:BBa_K199030:Experience

Designed by: Olivia Ho-Shing   Group: iGEM09_MoWestern_Davidson   (2009-07-11)

In order to have a visible expression of RFP or GFP, we constructed parts with RBS-RFP under the control of 4 different commonly-used promoters:

  • pBad (Potential induction with L-arabinose)
  • pLac (Potential induction with IPTG)
  • pLacIQ (Potential induction with IPTG)
  • pTet


We found that pLac induced with IPTG caused the greatest expression of RFP. This construct, pLac-RBS-RFP, was chosen as our control construct, representing 100% suppression of the engineered frameshift.

  • Notice the faint pink color with the pBad promoter only if Arabinose was added (tube number 3). We wanted to produce a part that gave a weak signal compared to all the strong promoter versions that existed.


Fluorescence readings were measured from biological duplicates (A and B) for each condition, and normalized to the cell density (RFP/OD). pBad constructs were induced with 2.5% L-arabinose. pLac and pLacIQ constructs were induced with 0.06% IPTG.



After 20 hours incubation, 400 ul of culture were pelleted down to visually compare colour. Left to right: 1. Negative 2. pBad 3. pBad+L-ara 4. pLac 5. pLacIQ 6. pLacIQ+IPTG 7. pTet 8. pLac+IPTG.




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