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Team:Sheffield/FadR
From 2014.igem.org
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| + | We investigated FadR expression using a Tecan Plate Reader. LB broth containing varying concentrations of oleic acid were innoculated with DH5a ''E. coli'' cells expressing fadR, fadR-RFP or consituative RFP. These were loaded into a clear flat bottomed black walled 96 well plate, some wells contained no cells and were used as blanks. Cells were grown in the Tecan plate reader overnight at 37oC with shaking for a period of 16 hours. OD600 measurements were taken every 30 minutes. In addition RFP readings were also taken with 565nm excitation and 605nm emission wavelengths. | ||
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| + | Values were normalised by OD and are shown as a mean of four repeats. | ||
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| + | The trend for all concentrations of oleic acid was for constitutive RFP to show an increase in expression (correlated to increased florescence) over time. In comparison there was no observable trend in fadR-RFP expressing cells. In addition there was no difference in the amount of fluorescence observed between the fadR-RFP and the fadR only and blank cells. Based on this information we suggest that the | ||
Revision as of 03:01, 18 October 2014
We investigated FadR expression using a Tecan Plate Reader. LB broth containing varying concentrations of oleic acid were innoculated with DH5a E. coli cells expressing fadR, fadR-RFP or consituative RFP. These were loaded into a clear flat bottomed black walled 96 well plate, some wells contained no cells and were used as blanks. Cells were grown in the Tecan plate reader overnight at 37oC with shaking for a period of 16 hours. OD600 measurements were taken every 30 minutes. In addition RFP readings were also taken with 565nm excitation and 605nm emission wavelengths.
Values were normalised by OD and are shown as a mean of four repeats.
The trend for all concentrations of oleic acid was for constitutive RFP to show an increase in expression (correlated to increased florescence) over time. In comparison there was no observable trend in fadR-RFP expressing cells. In addition there was no difference in the amount of fluorescence observed between the fadR-RFP and the fadR only and blank cells. Based on this information we suggest that the
