Difference between revisions of "Part:BBa K2379005"

Latest revision as of 03:20, 2 November 2017

RNA thermometer (FourU) with GFP under the regulation of pLAC

The RNA thermometer (BBa_K115002) is under the regulation of pLAC promoter (BBa_R0011). Following the RNA thermometer, the construct contains GFP(BBa_E0040) as the reporter which is followed by the double terminator (BBa_B0014). Expression of GFP is regulated by the RNA thermometer and hence will be produced only at 37 degree Celsius.

Usage and Biology

ABOUT THIS PART:

The part (BBa_K2379005) consists of RNA thermometer followed by GFP. The RNA thermometer(FourU) BBa_K115002(BBa_K115002) is a temperature sensitive element that shows higher expression of fluorescence at 37ºC and lower expression below 37ºC. The part works as expected and maximum level of fluorescence was observed at 37ºC.

CHARACTERIZATION STUDIES:

1.The above graph indicates the fluorescence expression at three different time periods (each recorded at an interval of 60 minutes.

2.The cell cultures were grown initially at temperature 37ºC until it reached an OD600 of 0.3. Then the cultures were transferred and incubated at three different temperatures: 32ºC, 37ºC and 42ºC.

3.Normalised fluorescence was calculated by dividing observed fluorescence by cell density at OD600, background fluoresence was minimized using a negative control.

4.Triplets were used for characterisation and the error bars indicate the Standard deviation of their mean values.

CONCLUSION:

The expression of GFP was found to be basal and minimum at 32ºC. A sudden induction in the expression was observed at the temperature 37ºC and it seemed to be gradually decreasing after 42ºC. This behaviour of RNA thermometer(FourU) observed supports the data given by other teams. Thus, the RNA thermometer(FourU) works as expected.

Sequence and Features