Part:BBa_K1172912

Part 1 of the Biosystem TetOR alive (pRha TetR alr)

Part 1 of the Biosystem M (pRha TetR alr)

Usage and Biology

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 1156
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1080
  Illegal BamHI site found at 1782
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 1198
  Illegal AgeI site found at 1498
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 955
  

Results

Characterization of the tetracycline operator TetO

Figure 12: Characterization of the bacterial growth of the TetO promoter with GFP (BBa_E0040) in the Biosafety-Strain K-12 ∆alr ∆dadX. The M9 media was supplemented with 5 mM D-alanine. It can be seen, that the bacteria grow faster on M9 minimal media glucose than on M9 minimal media glycerol.

Figure 13: Characterization of the fluorescence of TetO promoter with GFP (BBa_E0040) in the Biosafety-Strain K-12 ∆alr ∆dadX. The bacteria were cultivated on M9 minimal media with 5 mM D-alanine supplemented..

Figure 14: Specific growth rate of GFP behind the TetO promoter by the use of different carbon sources.

Characterization of the Biosafety-System TetOR alive

Figure 15: Characterization of the bacterial growth of the Biosafety-System on M9 minimal media glycerol. The figure compares the wild tpye K-12 and the Biosafety-Strain K-12 ∆alr ∆dadX and the induction by L-Rhamnose to pure glycerol.

Figure 16: Characterization of the fluorescence of the Biosafety-System TetOR alive. The figure compares the wild tpye k-12 and the Biosafety-Strain K-12 ∆alr ∆dadX and the induction by L-Rhamnose to pure glycerol.

Figure 17: Specific production rate, calculated by equation (1).

Conclusion of the Results

File:Team-Bielefeld-Biosafety-System-TetORalive-Overview.jpg

References

Saenger W. et al. (2000):Der Tetracyclin-Repressor – das Musterbeispiel für einen biologischen Schalter, In:[http://onlinelibrary.wiley.com/doi/10.1002/1521-3757%2820000616%29112:12%3C2122::AID-ANGE2122%3E3.0.CO;2-8/abstract Angewandte Chemie Volume 112, Issue 12, pages 2122–2133]