Difference between revisions of "Part:BBa K1913026"

 
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<partinfo>BBa_K1913026 short</partinfo>
 
<partinfo>BBa_K1913026 short</partinfo>
  
Wild type ptet-FixK2 hybrid promoter is the promoter that could not only be activated by light sensorsYF1-FixJ composite and but inhibited by TetR repressor. The sequence of upper control element and core element region are from wild type FixK2 hybrid promoter on genomic sequence of Bradyrhizobium japonicum. Additional tet operators are inserted in the core element region of promoter as second control element that results in transcription repression.  
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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 <i>Bradyrhizobium japonicum</i>. 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 (Fig.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.
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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.
  
 
https://static.igem.org/mediawiki/2016/9/9f/T--Wageningen_UR--light_result_26.jpg
 
https://static.igem.org/mediawiki/2016/9/9f/T--Wageningen_UR--light_result_26.jpg
  
Fig 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.
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<b>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.</b>
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
 
===Usage and Biology===
 
===Usage and Biology===

Latest revision as of 21:53, 19 October 2016


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]