RBS

Part:BBa_K3317067

Designed by: Delgadillo-Rubio Shakti   Group: iGEM19_UANL   (2019-10-20)


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.

Functional Parameters

[edit]
Categories
Parameters
None