Coding
mRFP1

Part:BBa_E2050:Experience

Designed by: ryu   Group: Antiquity   (2005-01-24)
Revision as of 23:08, 17 October 2014 by Dopeters (Talk | contribs) (→‎Applications of BBa_E2050)

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Applications of BBa_E2050

Part:BBa_E2050:Experience
Aberdeen iGEM 2010 Bio-brick Experience

The yeast mOrange open reading frame (Bio-brick E2050) was tested as part of a yeast expression construct, in which E2050 was translationally fused downstream of a tandem N-peptide open reading frame (Biobrick BBa_K385004). The gene fusion was placed under control of the GAL1 promoter (BioBrick K106699). The whole construct was cloned into a yeast shuttle vector and transformed into yeast. After growth of the transformant culture in medium containing galactose to induce the promoter, expression of mOrange was assessed using flow cytometry, (excitation wavelength 488nm, fluorescence wavelength 585nm). No mOrange fluorence was detectable by this method. However, a similar, related construct, in which the mOrange sequence was replaced by GFP, did express significant quantities of green fluorescent protein (see Aberdeen 2010 wiki for details ). This led us to conclude the mOrange sequence was non-functional in this particular expression construct.

iGEM14_Carnegie_Mellon. We characterized a set of fluorescent proteins consisting of BFP. GFP, YFP, OFP, and RFP. We calculated the signal-to-noise ratio of all the proteins in two different cell lines (MACH and Top10). mOrange (OFP) had a high signal-to-noise ratio in MACH cells and a lower signal-to-noise ratio in Top10 cells. In both cases it had a high signal-to-noise ratio relative to other fluorescent proteins. OFP was measured at (ex/em = 548nm/562nm).

591px-FP_MACH.png 591px-Top10_Best.jpg

User Reviews

UNIQ45f6f677163b2ed9-partinfo-00000001-QINU

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Slam

We were unable to detect mOrange fluorescence in our GAL1 regulated construct using this Bio-brick part. See above for more details.

Antiquity

This review comes from the old result system and indicates that this part did not work in some test.

UNIQ45f6f677163b2ed9-partinfo-00000004-QINU

User Reviews

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Tianjin iGEM

We used this part to confirm our yeast surface display system. Our result confirmed that this part is good. UNIQ45f6f677163b2ed9-partinfo-00000006-QINU