Difference between revisions of "Part:BBa K2378006"
Line 3: Line 3:
 
<partinfo>BBa_K2378006 short</partinfo>
 
<partinfo>BBa_K2378006 short</partinfo>
  
−
This is optimized coding sequence of PETase enzyme for <i>E. coli</i> BL21 with the addition of NhaR BBa_K1357002 (https://parts.igem.org/wiki/index.php?title=Part:BBa_K1357002) coding sequence. This part also use constitutive promoters BBa_J23106.
+
This is an optimized coding sequence of PETase enzyme for <i>E. coli</i> BL21 with the addition of NhaR BBa_K1357002 (https://parts.igem.org/wiki/index.php?title=Part:BBa_K1357002) coding sequence. NhaR enhances the transcription of pgaABCD operon which in turn induces the production of biofilm. This part also uses constitutive promoters BBa_J23106.  
 +
 
 +
<p style="font-size: 18px"><strong>Crystal Violet Biofilm Assay</strong></p>
 +
<p>Expression of NhaR induces rapid production of biofilm. In order to observe the effect of NhaR in this part, we tested the rate of biofilm production of bacteria transformed with this part containing NhaR compared to control bacteria without NhaR.</p>
 +
<br>
 +
<p>The experiment was done by growing bacterial cultures in a 96-well microtiter plate, and measuring its biofilm formation after 6, 12, 18, 24 hours of incubation. The complete protocol can be seen here (http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3182663/).</p>
 +
<br>
 +
<p>The result shows a more rapid biofilm production in cultures transformed with this part containing NhaR compared to its control without NhaR. </p>
 +
 
  
  

Revision as of 16:44, 1 November 2017


NhaR-PETase Constitutive Coding Device

This is an optimized coding sequence of PETase enzyme for E. coli BL21 with the addition of NhaR BBa_K1357002 (https://parts.igem.org/wiki/index.php?title=Part:BBa_K1357002) coding sequence. NhaR enhances the transcription of pgaABCD operon which in turn induces the production of biofilm. This part also uses constitutive promoters BBa_J23106.

Crystal Violet Biofilm Assay

Expression of NhaR induces rapid production of biofilm. In order to observe the effect of NhaR in this part, we tested the rate of biofilm production of bacteria transformed with this part containing NhaR compared to control bacteria without NhaR.


The experiment was done by growing bacterial cultures in a 96-well microtiter plate, and measuring its biofilm formation after 6, 12, 18, 24 hours of incubation. The complete protocol can be seen here (http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3182663/).


The result shows a more rapid biofilm production in cultures transformed with this part containing NhaR compared to its control without NhaR.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal NotI site found at 1002
    Illegal NotI site found at 1911
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1656
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 236