Part:BBa_K4193004:Design

SSRE promoter

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal XbaI site found at 239
  
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal XbaI site found at 239
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal XbaI site found at 239
  Illegal NgoMIV site found at 6
  Illegal NgoMIV site found at 46
  Illegal NgoMIV site found at 86
  Illegal NgoMIV site found at 126
  Illegal AgeI site found at 16
  Illegal AgeI site found at 56
  Illegal AgeI site found at 96
  Illegal AgeI site found at 136
  
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

Cytokinin isopentenyladenine (IP), as quorum sensing molecule, is the product of ATP catalyzed by AtlPT4. When the signal molecule IP reaches a certain concentration, it activates the SSRE promoter and increases the expression of production gene controlled by SSRE promoter in our experiment, to better characterize SSRE promoter and quorum sensing circuit, we put eGFP gene at the downstream of SSRE promoter.

Figure1.The signal pathway of IP-mediated quorum sensing

We successfully introduced cytokinin-mediated quorum sensing circuit into the genome of Aureobasidium melanogenum P16. To characterize this circuit and SSRE promoter, we added different concentration of IP and introduced eGFP at the downstream of SSRE promoter. We measured the fluorescence to figure out the dynamic regulation range.

Figure2.Gel electrophoresis results of P16 genome amplifying AtCRE1 gene (Lane1), transformed strain genome amplifying AtCRE1 (Lane2), P16 genome amplifying PTP2+eGFP (Lane3) and transformed strain genome amplifying PTP2+eGFP (Lane 4)

Source

synthesize from company

References

Yang, X., Liu, J., Zhang, J., Shen, Y., Qi, Q., Bao, X. and Hou, J. (2021). Quorum sensing-mediated protein degradation for dynamic metabolic pathway control in Saccharomyces cerevisiae. Metabolic Engineering, 64, pp.85–94. doi:10.1016/j.ymben.2021.01.010.

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