Difference between revisions of "Part:BBa K4245002"
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<partinfo>BBa_K4245002 short</partinfo> | <partinfo>BBa_K4245002 short</partinfo> | ||
| − | iSpinach aptamer | + | This part produces the fluorescent RNA aptamer iSpinach (BBa_K3380150) under the control of IPTG. iSpinach is a mutant of Spinach (BBa_K734002) developed by researchers at the University of Strasbourg. Compared to Spinach, iSpinach is less salt-sensitive, has higher thermal stability, and produces more fluorescence (Autour et. al, 2016). |
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| + | iSpinach is a fluorescent light-up aptamers (FLAP) that binds to 3’5-difluoro-4-hydroxybenzylidene imidazolinone (DFHBI), a small dye derived from the GFP fluorophore, to produce fluorescence (Paige et al., 2011). As shown in Figure 1, the aptamer and DFHBI bind together to produce green fluorescence, which has roughly 50% of the fluorescence intensity of enhanced GFP (Neubacher & Hennig, 2018). However, FLAPs can be more effective than GFP in biosensing as they bind to a fluorophore after transcription (RNA), while GFP requires additional translation for expression. Similar to other FLAPs, iSpinach is expressed within a transfer RNA (tRNA) scaffold, which shields the RNA from misfolding and degradation (Paige et al., 2011). | ||
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| + | This part works identically to BBa_K4245001, a composite part producing Spinach, and BBa_K4245003, a composite part producing iSpinach-D5-G30-A32; the only difference being the aptamer produced. This disparity allows accurate comparison of the basic aptamer parts. | ||
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| + | [[File:Characterization-figure-3.png|350px|link=]] | ||
| + | Figure 1. DFHBI and iSpinach aptamer binding to form RNA-fluorophore complex. | ||
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| + | The LacI protein represses the inducible promoter (BBa_R0010), which stops downstream transcription of the Spinach aptamer. When IPTG is present, LacI is inhibited, allowing for the transcription of the aptamer. Once DFHBI binds to the aptamer, the RNA-fluorophore complex produces a quantifiable green fluorescence. | ||
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<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here | ||
Revision as of 17:01, 6 October 2022
Status: 500
Content-type: text/html
Software error:
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For help, please send mail to this site's webmaster, giving this error message and the time and date of the error.
This part produces the fluorescent RNA aptamer iSpinach (BBa_K3380150) under the control of IPTG. iSpinach is a mutant of Spinach (BBa_K734002) developed by researchers at the University of Strasbourg. Compared to Spinach, iSpinach is less salt-sensitive, has higher thermal stability, and produces more fluorescence (Autour et. al, 2016).
iSpinach is a fluorescent light-up aptamers (FLAP) that binds to 3’5-difluoro-4-hydroxybenzylidene imidazolinone (DFHBI), a small dye derived from the GFP fluorophore, to produce fluorescence (Paige et al., 2011). As shown in Figure 1, the aptamer and DFHBI bind together to produce green fluorescence, which has roughly 50% of the fluorescence intensity of enhanced GFP (Neubacher & Hennig, 2018). However, FLAPs can be more effective than GFP in biosensing as they bind to a fluorophore after transcription (RNA), while GFP requires additional translation for expression. Similar to other FLAPs, iSpinach is expressed within a transfer RNA (tRNA) scaffold, which shields the RNA from misfolding and degradation (Paige et al., 2011).
This part works identically to BBa_K4245001, a composite part producing Spinach, and BBa_K4245003, a composite part producing iSpinach-D5-G30-A32; the only difference being the aptamer produced. This disparity allows accurate comparison of the basic aptamer parts.
Figure 1. DFHBI and iSpinach aptamer binding to form RNA-fluorophore complex.
The LacI protein represses the inducible promoter (BBa_R0010), which stops downstream transcription of the Spinach aptamer. When IPTG is present, LacI is inhibited, allowing for the transcription of the aptamer. Once DFHBI binds to the aptamer, the RNA-fluorophore complex produces a quantifiable green fluorescence.
Sequence and Features Status: 500 Content-type: text/html
Software error:
Global symbol "$groups" requires explicit package name at /websites/parts.igem.org/cgi/lib/User.pm line 84. Global symbol "$groups" requires explicit package name at /websites/parts.igem.org/cgi/lib/User.pm line 84. Global symbol "$groups" requires explicit package name at /websites/parts.igem.org/cgi/lib/User.pm line 85. Compilation failed in require at /websites/parts.igem.org/cgi/lib/IconBar.pm line 12. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/IconBar.pm line 12. Compilation failed in require at /websites/parts.igem.org/cgi/lib/BBWeb.pm line 9. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/BBWeb.pm line 9. Compilation failed in require at /websites/parts.igem.org/cgi/lib/Change.pm line 8. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/Change.pm line 8. Compilation failed in require at /websites/parts.igem.org/cgi/lib/Part.pm line 12. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/Part.pm line 12. Compilation failed in require at /websites/parts.igem.org/cgi/partsdb/putout.cgi line 8. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/partsdb/putout.cgi line 8.
For help, please send mail to this site's webmaster, giving this error message and the time and date of the error.