Difference between revisions of "Part:BBa K864100:Experience"
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| − | An improved version of the SYFP2 reporter with an N-terminal Cell-Penetrating peptide (CPP) signal was constructed by the UCopenhagen 2017 iGEM Team. The addition of the nona-argigine CPP tag was shown to give the SYFP2 biobrick the ability stain both <i>E. coli</i> and <i>P. putida</i> cells, presumably by causing internalisation. In addition the improved biobrick carries a C-terminal hexa-histidine tag. For more information please visit the improved part [[Part: | + | An improved version of the SYFP2 reporter with an N-terminal Cell-Penetrating peptide (CPP) signal was constructed by the UCopenhagen 2017 iGEM Team. The addition of the nona-argigine CPP tag was shown to give the SYFP2 biobrick the ability stain both <i>E. coli</i> and <i>P. putida</i> cells, presumably by causing internalisation. In addition the improved biobrick carries a C-terminal hexa-histidine tag. For more information please visit the improved part [[Part:BBa_K2455002|CPP-SYFP2]]. |
Revision as of 02:57, 2 November 2017
This experience page is provided so that any user may enter their experience using this part.
Please enter
how you used this part and how it worked out.
Applications of BBa_K864100
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). YFP had a very high signal-to-noise ratio in MACH cells and a low signal-to-noise ratio in Top10 cells relative to other fluorescent proteins. YFP was measured at (ex/em = 514nm/527nm).
User Reviews
UNIQf2abd5ec48884d7c-partinfo-00000000-QINU
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No review score entered. User:agynna |
iGEM Team Uppsala University 2012 This part was used as an fluorescent reporter in two ways by the Uppsala iGEM team 2012. It could easily be observed both in a Fluorescence activated cell sorter (FACS), on a UV table and even better on a Visi-Blue blue-light table. We recommend it whenever a strong fluorescent reporter is needed. 1. As a part of the sRNA screening system target construct (BBa_K864444), where it was a reporter of translational downregulation. Thanks to the very strong fluorescence, cells with very low levels of expression could easily be distinguished from non-fluorescent cells, using a FACS. In this experiment it was fused to a part of another protein, with the BBa_J18922 gly-ser linker in between. Fluorescence was not affected by this fusion. 2. In our promoter test, where it was coupled to different promoter as promoters-B0032-SYFP2 (using BBa_K864101). |
UNIQf2abd5ec48884d7c-partinfo-00000005-QINU
Improvement by ECUST 2017
The reporter sYPF2 has been codon-optimized for Rhodobacter sphaeroides 2.4.1 and it was successfully tested as a reporter in this chassis. We have made several measurements to show its fluorescence in Rhodobacter sphaeroides as well its properties (Fluorescence excitation/emission spectrum). For more information of the improved part, please go to the page of Part:BBa_K2308003.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Improvement by UCopenhagen 2017
An improved version of the SYFP2 reporter with an N-terminal Cell-Penetrating peptide (CPP) signal was constructed by the UCopenhagen 2017 iGEM Team. The addition of the nona-argigine CPP tag was shown to give the SYFP2 biobrick the ability stain both E. coli and P. putida cells, presumably by causing internalisation. In addition the improved biobrick carries a C-terminal hexa-histidine tag. For more information please visit the improved part CPP-SYFP2.