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GUS and GFP analyses

BZ Baozhen Zeng
TL Tianyu Li
WW Wei Wang
ZD Zhengrong Dai
JL Jie Li
ZX Zhiyuan Xi
KJ Kenan Jia
YX Yu Xing
BL Bingbing Li
JY Jiaqi Yan
WJ Wensuo Jia
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Approximately, 10 µL of the protein extract was mixed with 100 µL of reaction buffer (10 mM Tris-HCl (pH 8.0), 2 mM MgCl2, and 1 mM 4-MUG) in a 2-mL vial and incubated at 37 °C for 60 min, and 900 µL of 0.2 M Na2CO3 was then added to stop the reaction. The reaction mixture was subsequently transferred to a colorimetric cup adapted to a TBS-380 mini fluorometer with two excitation modes (UV; 365–395 nm and blue; 465–485 nm), which produced two emission spectra (440–470 nm for UV and 515–575 nm for blue). GUS activity was measured with the excitation set to the UV mode and with an emission wavelength of 365 nm. This activity is expressed as the relative fluorescence unit (RLU). For GFP measurement, the protein extract was transferred into a colorimetric cup, the excitation was set to blue, and the emission wavelength was 465 nm.

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