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Revision as of 16:16, 6 October 2022 by NickL (Talk | contribs)


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.
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This part produces the fluorescent RNA aptamer Spinach (BBa_K734002 BBa_K734002) under the control of IPTG. Originally generated by the Jaffrey Lab at Cornell University, the Spinach aptamer 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 (469 nm/501 nm) (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, Spinach is expressed within a transfer RNA (tRNA) scaffold, which shields the RNA from misfolding and degradation (Paige et al., 2011).


Figure 1. DFHBI and Spinach 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.

This part works identically to BBa_K4245002, a composite part producing iSpinach, 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.

                                   Characterization-figure-3.png
                                   Figure 3. Diagram of the BBa_K4245001 construct.

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.
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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.
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