Difference between revisions of "Part:BBa E0022"

 
 
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_E0022 short</partinfo>
 
<partinfo>BBa_E0022 short</partinfo>
  
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Cyan fluorescent protein (ECFP) reporter coding sequence without the Ribosome      Binding Site. Modified with an LVA tail for rapid degradation of the protein      and faster fall time for the emission. </P>
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Cyan fluorescent protein (ECFP) reporter coding sequence without the Ribosome      Binding Site. Modified with an LVA tail for rapid degradation of the protein      and faster fall time for the emission.  
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===Usage and Biology===
 
===Usage and Biology===
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Unspecified.
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_E0022 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_E0022 SequenceAndFeatures</partinfo>
  
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<!-- Uncomment this to enable Functional Parameter display
 
===Functional Parameters===
 
===Functional Parameters===
 
<partinfo>BBa_E0022 parameters</partinfo>
 
<partinfo>BBa_E0022 parameters</partinfo>
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===ECUST_China 2019 Charaterization===
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===Characterization===
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In ECUST_China 2019 characterization，we constructed a plasmid containing pLac and CI protein, adding BBa_E0022 eCFP as the reporter gene. We attempted to verify the function of this plasmid via observing the eCFP fluorescence by the comparison between the negative control and induced cells.
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<html><style>
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.exper-com-box{
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width: 80%;
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text-align: center;
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padding-bottom: 20px;
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}</style>
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<div class="exper-com-box"><img style="width:500px;" src="https://static.igem.org/mediawiki/parts/7/7f/T--ECUST_China--Figure.1_Fluorescence_intensity_of_eCFP.jpg"> <br><span style="font-size: 14px;">
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<b>Figure 1.</b> Fluorescence intensity of eCFP</span></div> </html>
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However, since the CI and eCFP was polycistron, both under the control of pLac, the expression of eCFP was disturbed by CI. The figure illustrated that as time went by, the expression of eCFP decreased, contrary to the prospective results. The characterization of eCFP in ECUST_China showed that BBa_E0022 was not suitable for the characterization of polycistron structure.
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==Functional Parameters: Austin_UTexas==
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<html>
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<body>
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<partinfo>BBa_E0022 parameters</partinfo>
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<h3><center>Burden Imposed by this Part:</center></h3>
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<figure>
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<div class = "center">
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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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</div>
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<figcaption><center><b>Burden Value: -0.4 ± 5.4% </b></center></figcaption>
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</figure>
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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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</body>
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</html>

Latest revision as of 02:41, 25 August 2020

enhanced cyan fluorescent protein derived from A. victoria GFP

Cyan fluorescent protein (ECFP) reporter coding sequence without the Ribosome Binding Site. Modified with an LVA tail for rapid degradation of the protein and faster fall time for the emission.


Usage and Biology

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]


ECUST_China 2019 Charaterization

Characterization

In ECUST_China 2019 characterization，we constructed a plasmid containing pLac and CI protein, adding BBa_E0022 eCFP as the reporter gene. We attempted to verify the function of this plasmid via observing the eCFP fluorescence by the comparison between the negative control and induced cells.



Figure 1. Fluorescence intensity of eCFP


However, since the CI and eCFP was polycistron, both under the control of pLac, the expression of eCFP was disturbed by CI. The figure illustrated that as time went by, the expression of eCFP decreased, contrary to the prospective results. The characterization of eCFP in ECUST_China showed that BBa_E0022 was not suitable for the characterization of polycistron structure.

Functional Parameters: Austin_UTexas

BBa_E0022 parameters

Burden Imposed by this Part:

Burden Value: -0.4 ± 5.4%

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.