Difference between revisions of "Part:BBa K775000"

 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K775000 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K775000 SequenceAndFeatures</partinfo>
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GFP intensity of S. cerevisiae cells transformed with ''FUS1pr-EGFP'' at t=5.10 after induction with alpha pheromone. Please also see our page: [http://2012.igem.org/Team:TU-Delft/part2 Reporter]
 
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[[Image:FUSGFP_Growth_Intensity_Div.png|960px]]
 
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GFP intensity and growth curves of S. cerevisiae transformed with ''FUS1pr-EGFP'' after induction with alpha pheromone
 
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[[Image:FUSGFP_Div_Time.png|960px]]
 
  
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===Functional Parameters===
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<img src="https://static.igem.org/mediawiki/2012/1/15/NR1_immuno.png" width="500"><br>
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<p>Immunolocalization of the receptor protein using FLAG treatment. A halo formation indicates appropriate localization in the membrane </p><br><br/>
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<img src="https://static.igem.org/mediawiki/igem.org/0/0d/DNAstainingNR1.jpg" width="500"><br>
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<p>Flow cytometry results of cells stained with Vybrant® DyeCycle™ Orange DNA Stain after 4.40 hours after activation with a saturative amount of nicotinic acid. In the middle and right picture a shift of peaks can be observed, corresponding with formation of more haploid cells (left) in comparison with diploid cells (right).
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==Functional Parameters: Austin_UTexas==
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<partinfo>BBa_K775000 parameters</partinfo>
 
<partinfo>BBa_K775000 parameters</partinfo>
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<h3><center>Burden Imposed by this Part:</center></h3>
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<center><img src = "https://static.igem.org/mediawiki/parts/f/fa/T--Austin_Utexas--no_burden_icon.png" style = "width:160px;height:120px"></center>
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<figcaption><center><b>Burden Value: 2.8 ± 6.2% </b></center></figcaption>
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<p> Burden is the percent reduction in the growth rate of <i>E. coli</i> cells transformed with a plasmid containing this BioBrick (± values are 95% confidence limits). This BioBrick did not exhibit a burden that was significantly greater than zero (i.e., it appears to have little to no impact on growth). Therefore, users can depend on this part to remain stable for many bacterial cell divisions and in large culture volumes. Refer to any one of the
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<a href="https://parts.igem.org/Part:BBa_K3174002">BBa_K3174002</a> - <a href="https://parts.igem.org/Part:BBa_K3174007">BBa_K3174007</a> pages for more information on the methods, an explanation of the sources of burden,  and other conclusions from a large-scale measurement project conducted by the <a href="http://2019.igem.org/Team:Austin_UTexas">2019 Austin_UTexas team</a>.</p>
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<p>This functional parameter was added by the <a href="https://2020.igem.org/Team:Austin_UTexas/Contribution">2020 Austin_UTexas team.</a></p>
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Latest revision as of 18:54, 3 September 2020

RI7-GPR109A Niacin chimeric GPCR

BBa_K775000 encodes a chimeric niacin-sensing G-protein coupled receptor which can be functionally expressed in S. cerevisiae. This chimeric receptor is composed of the ligand-binding domain of the mammalian GPR109A, flanked by the N- and C- terminals of the rat OR RI7.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 582
    Illegal BamHI site found at 113
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 478




Immunolocalization of the receptor protein using FLAG treatment. A halo formation indicates appropriate localization in the membrane




Flow cytometry results of cells stained with Vybrant® DyeCycle™ Orange DNA Stain after 4.40 hours after activation with a saturative amount of nicotinic acid. In the middle and right picture a shift of peaks can be observed, corresponding with formation of more haploid cells (left) in comparison with diploid cells (right).


Functional Parameters: Austin_UTexas

BBa_K775000 parameters

Burden Imposed by this Part:

Burden Value: 2.8 ± 6.2%

Burden is the percent reduction in the growth rate of E. coli cells transformed with a plasmid containing this BioBrick (± values are 95% confidence limits). This BioBrick did not exhibit a burden that was significantly greater than zero (i.e., it appears to have little to no impact on growth). Therefore, users can depend on this part to remain stable for many bacterial cell divisions and in large culture volumes. Refer to any one of the BBa_K3174002 - BBa_K3174007 pages for more information on the methods, an explanation of the sources of burden, and other conclusions from a large-scale measurement project conducted by the 2019 Austin_UTexas team.

This functional parameter was added by the 2020 Austin_UTexas team.