Difference between revisions of "Part:BBa K864100:Experience"

Latest revision as of 03:00, 2 November 2017

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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

UNIQf26040d4c8a434a7-partinfo-00000000-QINU

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).

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UNIQf26040d4c8a434a7-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

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

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 <partinfo>BBa_K2308003  parameters</partinfo>
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-===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 <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]].