Part:BBa_K864400:Experience
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how you used this part and how it worked out.
Applications of BBa_K864400
Team INSA Lyon 2016
Author: Gonthier Mathilde
Summary: The INSA Lyon 2016 team used this part in order to characterize it. A GFP or a CFP coding genes were set after this promotor, by measuring the fluorescence of the expressions cultures having these constructs the team was able to gather some informations about the behavior of this promotor. For further informations you may consult both BBa_K1934040 and BBa_K1934050 registry pages.
Team Uni Padua IT
We used pTAC as a basic part in an expression cassette combining RBS (BBa_B003) and the coding sequence that encodes a carrier protein and the Dehalogenase type II gene (BBa_K5109016)
We tested the expression in TOP10 F’ Escherichia coli cells, since they keep the protein expression constantly repressed in basic conditions. We then performed growth test of the transformed cells containing the expression cassette BBa_K5109023 compared to the growth of wild type TOP10 F’ cells.
Cellular growth was measured using a plate reader by taking the OD600 value every five minutes for 14 hours.
We then used the matrix of data generated by the plate reader to create the growth curves and analyze how they changed in the different concentrations of IPTG: respectively, 0.5 uM, 5uM, 50uM and 500 uM IPTG.
Overall, cellular growth was lower under IPTG induction compared to wild type colonies.
When analysing the data obtained from the growth rates for DeHa2, increasing the concentration of IPTG leads to a slow but constant decrease in the growth of the colonies. Further investigation is needed to decide the best IPTG concentration suitable in order to use our composite part.
User Reviews
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mbecich |
This part function as described by our team, it's an efficient promotor. But notice that it tends to leak after a few hours. |
UNIQ43e6bef17896c4ec-partinfo-00000005-QINU