For numerical value evaluation, cultured cells were sample in every two hours in 96 well plates. The plates were black-colored and flat bottom plates, and measured by plate reader.(TECAN Infinite M200 in Bio Molecular Engineering Laboratory in KAIST, Republic of Korea) The GFPmut3b and mRFP are have maximum excitation wavelength at 501nm and 584nm respectively. Those of emission are 511nm and 607nm. However, the intensity of GFP was measured with excitation wavelength of 501nm and 536nm in emission and 584nm and 619nm in case of mRFP. Because, if the excitation and emission wavelength are too close, the signal interferes and peaks are somewhat unclear.
(Left-A) Induced at 6hr, GFP intensity of double transformant pFlipFlopGFP-pFlipFlopIntergrase(purple)decreased, while the uninduced cells (yellow green) still express GFP. Means that Bxb1 integrase inverts pFlipFlopGFP into RFP state.
(Left-B) Induced at 6hr, the increment of RFP intensity of double transformant pFlipFlopGFP-pFlipFlopIntergrase(purple) is relatively larger than the uninduced group (yellow green). Means that Bxb1 integrase inverts pFlipFlopGFP into RFP state.
(Right-A) Induced at 6hr, the increment of GFP intensity of double transformant pFlipFlopRFP-pFlipFlopIntergrase(purple) is relatively larger than the uninduced group (yellow green). Means that Bxb1 integrase inverts pFlipFlopRFP into GFP state.
(Right-B) Induced at 6hr, the increment of RFP intensity of double transformant pFlipFlopRFP-pFlipFlopIntergrase(purple) is relatively smaller than uninduced group (yellow green). Means that Bxb1 integrase inverts pFlipFlopRFP into GFP state.
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