Reporter

Part:BBa_K4244063

Designed by: Arthur Vancolen   Group: iGEM22_Wageningen_UR   (2022-10-09)
Revision as of 08:48, 12 October 2022 by ArthurVancolen (Talk | contribs)


Constitutive expression of sfGFP with HlyA secretion tag

sfGFP BBa_K4244004 was fused to HlyA BBa_K554002 (fused at the C-terminus) to secrete sfGFP and use it as an extracellular reporter. It is under an constitutive medium strength promoter BBa_K4244057.

Secretion assay

Figure 1: Time series measurement of total, pellet and supernatant fraction of sfGFP under the HlyA signal peptide expressed with constitutive promoter. Measurement was performed every two hours for eight hours. No significant fluorescence was detected in the supernatant fraction.

Discussion

No secretion was observed in E. coli Nissle. This may be due to the too high expression level of sfGFP under BBa_K4244057. This experiment still need adjustment and fine tuning to prove the secretion of sfGFP.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None