Composite

Part:BBa_K4340605

Designed by: Chein Yueh Liu   Group: iGEM22_PuiChing_Macau   (2022-09-29)


sfGFP

sfGFP(BBa_K2762014) is able to show fluorescence in an acidic environment. We use this plasmid to test the growth curve in our hydroponic system.

E.coli (sfGFP_pET11a) growth curve

These are the results of our sfGFP_pET11a test. We measured the pH, OD 600, and fluorescence of the cell culture of the transformed E.coli. We use these three indicators to validate the pH neutralizing function and the survival and growth curve of the E.coli transformed with sfGFP_pET11a plasmid.

1. Fluorescence test

Figure 1. The Fluorescence Rate in pH 5 initial environment of sfGFP_pET11a and Pasr-glsA_pET11a.
Figure 2. The Fluorescence Rate in pH 7 initial environment of sfGFP_pET11a and Pasr-glsA_pET11a.
Figure 3. The Fluorescence Rate in pH 9 initial environment of sfGFP_pET11a and Pasr-glsA_pET11a.


The fluorescence of sfGFP in pH 5 is significantly higher than the fluorescence of Pasr-glsA. It is possibly because the protein size of the sfGFP is smaller than Pasr-glsA, which at the same time produces ammonia to neutralize the environment. In a pH 7 environment, the fluorescence of both sfGFP and Pasr-glsA is relatively similar and reached the same point at the eighth hour. In a pH 9 environment, both fluorescence of sfGFP and Pasr-glsA are low compared to data in pH 5 and 7. This proved that sfGFP and Pasr-glsA, which have the same acid promoter (asr) show low fluorescence in a high-pH environment.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 1283
    Illegal PstI site found at 5809
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1238
    Illegal PstI site found at 5809
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1349
    Illegal BamHI site found at 1205
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 1283
    Illegal PstI site found at 5809
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 1283
    Illegal PstI site found at 5809
    Illegal NgoMIV site found at 1381
    Illegal NgoMIV site found at 2969
    Illegal NgoMIV site found at 3129
    Illegal NgoMIV site found at 3483
    Illegal AgeI site found at 752
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 5635
    Illegal SapI site found at 4552


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