Difference between revisions of "Part:BBa K2379009"

Revision as of 02:49, 2 November 2017

Adaptor with constitutive RBS

This construct consists of the adaptor BBa_K2379008(BBa_K2379008) and is under the influence of the pLac promoter(BBa_R0011). In addition to the adaptor, a constitutive RBS (BBa_B0034) is used to regulate the transcription of the genes downstream. Following the adaptor is the reporter protein GFP (BBa_E0040) and the double terminator(BBa_B0014). Since it is regulated by a constitutive RBS, the circuit will be active at all times.

Usage and Biology

ABOUT THIS PART :

This part(BBa_K2379009)is an variant of adaptor which is regulated by constitutive RBS(BBa_B0034) placed downstream of pLac promoter(BBa_R0011). Since this part contains constitutive RBS, it shows similar levels of fluorescence expression irrespective of temperatures. The part works as expected and when GFP is placed downstream of this gene, similar levels of fluorescence was observed at 32ºC, 37ºC and 42ºC.

CHARACTERISATION STUDIES FOR THIS PART:

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 32ºC 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.

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

CONCLUSION:

The part works as expected. Since there is no temperature based regulatory element, similar fluorescence was observed at all the three different temperatures irrespective of the time points.

Sequence and Features