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

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===Applications of BBa_K864100===
 
===Applications of BBa_K864100===
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'''iGEM14_Carnegie_Mellon'''.
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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).
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https://static.igem.org/mediawiki/2014/thumb/c/c1/FP_MACH.png/591px-FP_MACH.png
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https://static.igem.org/mediawiki/2014/thumb/9/9f/Top10_Best.jpg/591px-Top10_Best.jpg
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=== User Reviews===
 
=== User Reviews===
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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.  
 
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.  
  
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1. As a part of the sRNA screening system target construct (<partinfo>K864444</partinfo>), 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.  
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1. As a part of the sRNA screening system target construct (<partinfo>K864444</partinfo>), 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 <partinfo>J18922</partinfo> gly-ser linker in between. Fluorescence was not affected by this fusion.  
  
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2. In our promoter test, where it was coupled to different promoter as promoter-B0032-SYFP2 (using <partinfo>K864101</partinfo>).  
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2. In our promoter test, where it was coupled to different promoter as promoters-B0032-SYFP2 (using <partinfo>K864101</partinfo>).  
 
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<!-- DON'T DELETE --><partinfo>BBa_K864100 EndReviews</partinfo>
 
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===Improvement by ECUST 2017===
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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]].
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K2308003 SequenceAndFeatures</partinfo>
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===Functional Parameters===
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<partinfo>BBa_K2308003  parameters</partinfo>
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Latest revision as of 03:00, 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).

591px-FP_MACH.png 591px-Top10_Best.jpg

User Reviews

UNIQ9659f06f33087aca-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).

;


UNIQ9659f06f33087aca-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]