Difference between revisions of "Team:KAIT Japan/Results"

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(The fluorescent lamp is including some wavelength. However, 400 nm of energy was stronger than 500 nm of energy. So, Dronpa fluorescent by fluorescent lamp.)<br>
 
(The fluorescent lamp is including some wavelength. However, 400 nm of energy was stronger than 500 nm of energy. So, Dronpa fluorescent by fluorescent lamp.)<br>
 
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<img src="https://static.igem.org/mediawiki/2015/a/af/%E3%82%AD%E3%83%A3%E3%83%97kekka4.PNG"><br>
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We irradiated 400 nm (5 hours) and 500 nm (5 hours) collectively for Dronpa 145K and 145N.<br>
 
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Revision as of 04:02, 18 September 2015

Results


[Expression of Dronpa 145K and Dronpa 145N]
We were cloning Dronpa 145K and Dronpa 145N in E.coli. And we expressed these parts using pcold vector.



E.coli fluorescence when we irradiated about 400 nm.



[Purification of Dronpa 145N and 145K]
We purified Dronpa 145N and Dronpa 145K by His-tag purification.



We irradiated about 400 nm at purification of Dronpa 145N. It fluorescence green.



[SDS-PAGE]
We expressed Dronpa 145N, 145K and luciferase fusion protein at pcold vector in E.coli and obtained these proteins at His-tag purification.



Dronpa is about 28.8 kDa. The 1 lane is the molecular maker. 2 lane and 3 lane are respectively Dronpa 145K and 145N.
We were obtained Dronpa 145N and 145K.



[Fluorescence intensity]
We analyzed for fluorescence intensity of Dronpa 145N and Dronpa 145K when we irradiated 400 nm or 500 nm.
We were irradiated 500 nm by Figure4 of machine. This is 68mW and it can irradiate LED light. This machine can choose wavelength by combining wavelength of 3 kinds. This machine spectrum is Figure5. However, we don’t have machine which isn't able to irradiate 400 nm. So we irradiated 400 nm using light of the fluorescent lamp. This is 32W. Spectrum of fluorescent lamp is Figure6.
(The fluorescent lamp is including some wavelength. However, 400 nm of energy was stronger than 500 nm of energy. So, Dronpa fluorescent by fluorescent lamp.)



We irradiated 400 nm (5 hours) and 500 nm (5 hours) collectively for Dronpa 145K and 145N.