Team:Carnegie Mellon/mRFP1

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Latest revision as of 01:21, 28 September 2013

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)

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-<h2>mRFP1 Characterization</h2>
+<h2>mRFP1 characterization</h2>
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-[[Image:CRQ-mRFP1.png|thumb|600px|center|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.]]
+[[Image:CRQ-mRFP1.png|thumb|300px|center|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.]]
 <html>
 <p align="center"><b> Characterization of the Photostability of mRFP1</b></p></html>
 [[Image:MRFP1.png|center|thumb|400px|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. <html><br />
-<table cellpadding="2" border="1px" cellspacing="0" align="center" width="70%">
+<table cellpadding="2" border="1px" cellspacing="0" align="center" width="40%">
 <caption><p align="justify"><b>Table 3</b> Tecan Safire II Parameters</p></caption>
 <tr><td><b>Excitation (nm)</b></td><td>585</td></tr>
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 </table>
 <br />
-<table cellpadding="2" border="1px" cellspacing="0" align="center" width="50%">
+<table cellpadding="2" border="1px" cellspacing="0" align="center" width="40%">
 <caption><p align="justify"><b>Table 4</b> Shows the fluorescence data over time during photobleaching.</p></caption>
 <tr><td align="center"><b>Time (minutes)</b></td><td align="center"><b>Fluorescence (RFU)</b></td>
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 <br>
 <h2>References</h2>
+ Campbell,R.E., Tour,O., Palmer,A.E., Steinbach,P.A., Baird,G.S., Zacharias,D.A. and Tsien,R.Y. <i>A monomeric red fluorescent protein</i>. Proc. Natl. Acad. Sci. U.S.A. 99 (12), 7877-7882 (2002)
 </html>