Team:Carnegie Mellon/mRFP1

From 2013.igem.org

Killer Red

mRFP1 characterization

The QYG-derived chromophore of the mRFP1 protein. (Z)-2-(2-(4-amino-1-imino-4-oxobutyl)-4-(4-hydroxybenzylidene)-5-oxo-4,5-dihydro-1H-imidazol-1-yl)acetic acid. The extended π-system is highlighted in red.

Characterization of the Photostability of mRFP1

Photobleaching curve of mRFP1 with a HBO100 mercury-arc lamp

XL10 Ultracompetent cells were transformed with <partinfo>BBa_E1010</partinfo> cloned with <partinfo>BBa_B0034</partinfo> as the RBS and <partinfo>BBa_R0010</partinfo> as the wild-type lac promoter and induced overnight with IPTG.The overnight was bleached for 180 minutes with HBO100 (100W Mercury-arc lamp). Fluorescence data was taken using a Tecan Safire II with the parameters shown in Table 3. Fluorescence values are shown in Table 4.

**Table 3** Tecan Safire II Parameters
Excitation (nm)	585
Emission (nm)	610
Excitation bandwidth (nm)	10
Emission bandwidth (nm)	10
Gain	63
Number of reads	10
Integration Time (microseconds)	40

**Table 4** Shows the fluorescence data over time during photobleaching.
Time (minutes)	Fluorescence (RFU)
0	48694
20	43083
40	36842
60	30239
80	31281
100	25273
120	21467
140	18081
160	15251
180	14427

References

Campbell,R.E., Tour,O., Palmer,A.E., Steinbach,P.A., Baird,G.S., Zacharias,D.A. and Tsien,R.Y. A monomeric red fluorescent protein. Proc. Natl. Acad. Sci. U.S.A. 99 (12), 7877-7882 (2002)