Part:BBa_K2447000
Extracellular phosphate sensor with GFP reporter
Improvement over previous iGEM part BBa_K116404 (NYMU Taipei 2008)
The two parts (Bba_K2447000 & BBa_K116404) listed below are inserted into pBbE2k backbones and subsequently characterised in E.coli MG 1655. These cells were grown in LB medium before transferring into MOPS medium (a minimal nutrient medium) for characterization with a micro-plate reader. Varying concentrations of phosphate ions from 0 to 1000 uM were added and GFP productions were monitored.
By replacing the weaker RBS 32 (as utilised by the Taiwanese team) with a stronger binding affinity RBS 34 as proposed by iGEM NUS team 2017, we have elucidated much stronger GFP expression and also improved the sensitivity of the part to varying concentrations of phosphate ions. Previously, the Taiwanese part [http://2008.igem.org/Team:NYMU-Taipei/Project/Phosphate had reported]much lower levels of GFP productions for all phosphate concentrations. Our improved construct had yielded on average 40 fold increase (Figure 2) in GFP expression when compared to the original construct. In addition, our proposed phosphate sensor is more sensitive (Figure 3) at higher levels of phosphate concentrations, and able to yield different levels of repressed GFP productions. On the other hand, the Taiwanese construct remains not sensitive to high amount of phosphate concentrations, yielding similar level of repressed GFP productions (Figure 4) when the phosphate concentrations is above 50 uM.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1162
chassis | E.coli (K12) |