Difference between revisions of "Part:BBa E2050:Experience"
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We were unable to detect mOrange fluorescence in our GAL1 regulated construct using this Bio-brick part. See above for more details. | We were unable to detect mOrange fluorescence in our GAL1 regulated construct using this Bio-brick part. See above for more details. | ||
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Revision as of 21:14, 6 November 2010
This experience page is provided so that any user may enter their experience using this part.
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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.
User Reviews
UNIQ31eb90822e1c77eb-partinfo-00000001-QINU
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We were unable to detect mOrange fluorescence in our GAL1 regulated construct using this Bio-brick part. See above for more details. UNIQ31eb90822e1c77eb-partinfo-00000003-QINU User ReviewsUNIQ31eb90822e1c77eb-partinfo-00000004-QINU
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