Regulatory

Part:BBa_K1913026

Designed by: Tianhe Wang   Group: iGEM16_Wageningen_UR   (2016-10-14)


Wild type ptet-FixK2 hybrid promoter

Wild type ptet-FixK2 hybrid promoter is a promoter which can not only be activated by the light sensor YF1-FixJ but also inhibited by the TetR repressor. The sequences of the upper control element and core element region are from wild type FixK2 promoter on genomic sequence of Bradyrhizobium japonicum. Additional tet operators are inserted in the core element region of the promoter as second control elements which result in transcription repression.

In order to test the hybrid promoters, we constructed five composite parts with mRFP gene as reporter (BBa_K19103027, BBa_K19103028, BBa_K19103029, BBa_K19103030, BBa_K19103031). These composite parts were co-transformed with the light sensor part (BBa_K19103034) into E.coli strain BL21, cultured under dark for 24h and their fluorescence was tested. We included as controls cells that only contained the Fixk2 composite parts. The results (Figure 1) illustrated that hybrid promoter BBa_K1913026 has only little different compared to control,which suggested that this promoter is not vert sensitive one for being induces by FixJ.

T--Wageningen_UR--light_result_26.jpg

Figure 1: Ratio of fluorescence value and absorbance of Fixk2 composite. Fixk2 composite and control was cultured under dark condition over 24h. Emission and excitation wavelength of mRFP are 607 and 584 nm respectively. 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]


[edit]
Categories
Parameters
None