Difference between revisions of "Part:BBa K3900021"
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The improved pH stability was determined by measuring the relative fluorescence of mRuby3 and mCRISPRed at different pH conditions. The fluorescence proteins were expressed in <I>Escherichia coli</I> BL21 DE3 and purified via IMAC beforehand.
 
The improved pH stability was determined by measuring the relative fluorescence of mRuby3 and mCRISPRed at different pH conditions. The fluorescence proteins were expressed in <I>Escherichia coli</I> BL21 DE3 and purified via IMAC beforehand.
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After purification, both protein eluates were rebuffered and concentrated to a volume of 1.5 mL. The concentration was measured by bradford assay and a concentration of 81.6 μmol/L for mRuby3 and 20.4 μmol/L for mCRISPRed was determined. The samples were diluted to the same concentration, before fluorescence measurement at different pH conditions using the TECAN plate reader.

Revision as of 06:12, 20 October 2021


emission592 nm
excitation460 nm

mCRISPRed - improvement of mRuby3

This part is an improved version of the red fluorescent protein mRuby3 (BBa_M50009), that displays an enhanced stability at low pH conditions. The original part can be found at: https://parts.igem.org/Part:BBa_M50009

Measurement
The improved pH stability was determined by measuring the relative fluorescence of mRuby3 and mCRISPRed at different pH conditions. The fluorescence proteins were expressed in Escherichia coli BL21 DE3 and purified via IMAC beforehand.
After purification, both protein eluates were rebuffered and concentrated to a volume of 1.5 mL. The concentration was measured by bradford assay and a concentration of 81.6 μmol/L for mRuby3 and 20.4 μmol/L for mCRISPRed was determined. The samples were diluted to the same concentration, before fluorescence measurement at different pH conditions using the TECAN plate reader.