Part:BBa_K3317067

37 � C inducible RNA thermometer

RNA thermometer, by subjecting the cells to temperatures greater than 37 ℃ the expression of the proteins placed downstream is allowed, this sequence is an improvement of (BBa_K1140006)

Usage and Biology

The primary objective was to perform an in silico design to reduce the range of operation of previously reported thermometers (28-37° C), obtaining a more precise range and allowing greater control of the system.

Tests were performed in conjunction with the (BBa_K1140006) thermometer from 25 to 40° C, measuring at 586/610 nm (excitation/emission for mCherry) and 660 nm to obtain optical density of the culture and normalize results, which showed that the new thermometer controlled the expression of the reporter in a more precise range, like the predicted theoretically. The in silico design allows the optimization of regulatory elements, such as RNA thermometers, reducing the number of experiments to a minimum.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 63
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

BBa_K1140006 was redesigned in the area of ​​the U6 thermometer, in order to reduce the operation temperature (28-37° C). The new piece, BBa_K3317067, showed as RNA Thermometer Pro in the features, consists of 121 nucleotides, with a Hairpin formation in the position 91 to 101, before the RBS (AGGAGA), allowing greater control of the system.