Difference between revisions of "Part:BBa K2043014"

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<partinfo>BBa_K2043014 short</partinfo>
 
<partinfo>BBa_K2043014 short</partinfo>
  
GFP-FBD7
 
  
<!-- Add more about the biology of this part here
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In the context of Paris Bettencourt 2016 iGEM project, Frank&Stain, we looked for fabric binding domains with phage display in order to optimise our enzymes on the topic of pigment degradation. More information can be consulted in the wiki, but we wanted to degrade wine stains, and we developed the FBD in order to direct the enzymatic activity to the fabric. <br><br>
===Usage and Biology===
+
 
 +
We fused the GFP gene with the FBD7 (in the N-terminal of the GFP) in order to understand two things: (i)if the GFP fluorescence would be affect by the binding of the FBD and (ii)if the binding capacity would be affected. <br><br>
 +
 
 +
The results for FBD7-GFP are as follows:
 +
https://static.igem.org/mediawiki/parts/5/56/Paris_Bettencourt-biobricks_gfp7.png
 +
These experiments were performed by obtaining cell extract of cells of a strain of <i>E. coli<(i> overexpressing FBD7-GFP, incubating the cell extract with the fabrics overnight, and then performing 2 washes  with the indicated solutions.
 +
<br><br>
  
 
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<h1>GFP-FBD7</h1>
 
<h1>GFP-FBD7</h1>
  
<pre>
+
Jain, P., Soshee, A., Narayanan, S. S., Sharma, J., Girard, C., Dujardin, E., & Nizak, C. (2014). Selection of arginine-rich anti-gold antibodies engineered for plasmonic colloid self-assembly. The Journal of Physical Chemistry C, 118(26), 14502-14510.<br><br>
 +
 
 +
Soshee, A., Zürcher, S., Spencer, N. D., Halperin, A., & Nizak, C. (2013). General in vitro method to analyze the interactions of synthetic polymers with human antibody repertoires. Biomacromolecules, 15(1), 113-121.<br><br>
  
                              ,oooooo888888888oooooo.
+
Boyer, S., Biswas, D., Soshee, A. K., Scaramozzino, N., Nizak, C., & Rivoire, O. (2016). Hierarchy and extremes in selections from pools of randomized proteins. Proceedings of the National Academy of Sciences, 201517813.<br><br>
                        .oo88^^^^^^            ^^^^^Y8o.
+
                      .dP'                              `Yb.
+
                    dP'                                  `Yb
+
                  .dP'                                    `Yb.
+
                dP'                                        `Yb
+
                d8                                            8b
+
              ,8P                                            `8b
+
              88'                                              88
+
              88                                              88
+
              dP                                              88
+
              d8'                                              88
+
              8P                                              ,dY
+
            ,dP                                                88'
+
          CP  ,,.....                ,,.....                88
+
          `b,d8P'^^^'Y8b          ,d8P'^^^'Y8b.             ,dY
+
            dP'        `Yb        dP'        `Yb            88'
+
          dP            Yb      dP            Yb          88
+
          dP    db        Yb    dP    db        Yb        ,dY
+
          88    YP        88    88    YP        88        88'
+
          Yb              dP    Yb              dP        88
+
          Yb            dP      Yb            dP        ,dY
+
          dP`Yb.       ,dP'        `Yb.      ,dP'          88'
+
        CCo_ `YbooooodP'            `YbooooodP'            88
+
          dP"oo_    ,dP            Ybo__                    88
+
        88    "ooodP'                ""88oooooP'          88
+
          Yb .ood""                                        ,dY
+
          ,dP"                                            88'
+
        ,dP'                                              88
+
      dP'    ,dP'    ,dP      ,dP'      .bmw.           88
+
      d8    dP      dP        dP        o88888b          88
+
      88    dP      dP      ,dP      o8888888P          88
+
      Y8.   88      88      d8P      o8888888P          ,dY
+
      `8b  Yb      88      88      ,8888888P          88'
+
      88    Yb    Y8.      88      888888P'            88
+
      88    `8b    `8b      88      88                ,dY
+
      88    88    88      Yb.      Yb                88'
+
      Y8.  ,Y8    ,Y8      ,88      ,8b                88
+
        `"ooo"`"oooo" `"ooooo" `8boooooP                ,8Y
+
            88boo__      """      """  ____oooooooo888888
+
          dP  ^^""ooooooooo..oooooo"""^^^^^            88
+
          88              88                          88
+
          88              88                          88
+
          Yboooo__        88          ____oooooooo88888P
+
            ^^^"""ooooooooo''oooooo"""^^^^^
+
</pre>
+

Revision as of 11:22, 24 October 2016


GFP-FBD7


In the context of Paris Bettencourt 2016 iGEM project, Frank&Stain, we looked for fabric binding domains with phage display in order to optimise our enzymes on the topic of pigment degradation. More information can be consulted in the wiki, but we wanted to degrade wine stains, and we developed the FBD in order to direct the enzymatic activity to the fabric.

We fused the GFP gene with the FBD7 (in the N-terminal of the GFP) in order to understand two things: (i)if the GFP fluorescence would be affect by the binding of the FBD and (ii)if the binding capacity would be affected.

The results for FBD7-GFP are as follows: Paris_Bettencourt-biobricks_gfp7.png These experiments were performed by obtaining cell extract of cells of a strain of E. coli<(i> overexpressing FBD7-GFP, incubating the cell extract with the fabrics overnight, and then performing 2 washes with the indicated solutions.

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]



GFP-FBD7

Jain, P., Soshee, A., Narayanan, S. S., Sharma, J., Girard, C., Dujardin, E., & Nizak, C. (2014). Selection of arginine-rich anti-gold antibodies engineered for plasmonic colloid self-assembly. The Journal of Physical Chemistry C, 118(26), 14502-14510.

Soshee, A., Zürcher, S., Spencer, N. D., Halperin, A., & Nizak, C. (2013). General in vitro method to analyze the interactions of synthetic polymers with human antibody repertoires. Biomacromolecules, 15(1), 113-121.

Boyer, S., Biswas, D., Soshee, A. K., Scaramozzino, N., Nizak, C., & Rivoire, O. (2016). Hierarchy and extremes in selections from pools of randomized proteins. Proceedings of the National Academy of Sciences, 201517813.