Part:BBa_K1033916:Experience

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UNIQ5027b59e073c14d5-partinfo-00000000-QINU UNIQ5027b59e073c14d5-partinfo-00000001-QINU

••••• Hong Kong-CUHK iGEM 2017

Fluorescent properties of amajLime Instead of chromorptein, we proposed that amajLime is a fluorescent protein with the property of showing pigmentation, like BBa_E1010. We charaterised spectral properties of amajLime and found its max excitation and emission wavelength at 445 nm nd 485 nm respectively. Fig.1 Emission and Excitation spectra Charaterization of pH stability of amajLime We transformed part BBa_K1033916 with constituitive promoter: J23100 in C41 and grew in 2XYT for 24 hours. Purifying the amajLime by Ion Exchange Chromatography and Hydrophobic Interaction Chromatography, we measured the fluoresece of purified amajLime, which is diluted to 10µg/100µl (total 200µl) in triplicates, in different buffers (ranges from pH2 to pH12). The result shows that the stability drops dramatically in condition below 6 and attains maxima fluorescence at pH 8. Fig.2 Vary pH attributed to different fluorescent intensity of RFP Table 1 Plate reader setting of fluorescent measurement Basic Settings Measurement Type Fluorescence Microplate name COSTAR 96 Scan mode orbital averaging Scan diameter [nm] 3 Excitation 470-15 Emission 515-20 Dichronic filter auto 491.2 Gain 500 Focal height [nm] 9

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