Difference between revisions of "Team:BJRS China/Improve"

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<p><font size="4">To characterize the surface display efficiency of INP better, we added a strong promoter <html><a href='http://parts.igem.org/Part:BBa_J23104'>BBa_J23104</a></html> and RBS <html><a href='http://parts.igem.org/Part:BBa_B0032'>BBa_B0032</a></html> to the upstream of INP, and changed the YFP to sfGFP, to construct our parts <html><a href='http://parts.igem.org/Part:BBa_K2833009'>BBa_K2833009</a></html>(Fig.1)</font></p>
 
<p><font size="4">To characterize the surface display efficiency of INP better, we added a strong promoter <html><a href='http://parts.igem.org/Part:BBa_J23104'>BBa_J23104</a></html> and RBS <html><a href='http://parts.igem.org/Part:BBa_B0032'>BBa_B0032</a></html> to the upstream of INP, and changed the YFP to sfGFP, to construct our parts <html><a href='http://parts.igem.org/Part:BBa_K2833009'>BBa_K2833009</a></html>(Fig.1)</font></p>
  
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                 <img src="https://static.igem.org/mediawiki/parts/b/b5/T--BJRS_China--improvement009.png"width="50%" >
 
                 <img src="https://static.igem.org/mediawiki/parts/b/b5/T--BJRS_China--improvement009.png"width="50%" >
 
                 <figcaption><div style="padding-left:300px;padding-right:300px"><b>Figure 1</b> the framework of BBa_K2833009.
 
                 <figcaption><div style="padding-left:300px;padding-right:300px"><b>Figure 1</b> the framework of BBa_K2833009.

Revision as of 02:30, 15 October 2018

BJRS


Improvement

Design

Our project used surface display system, and we choose the INP-YFP expressiong part BBa_K523013 built by Edinburgh 2011 to construct the INP-mediated surface display for our interest protein. While testing the surface display efficiency of this part, we found that the fluorescence intensity was too weak both under the uv-light（adjusted to the same OD600）(Fig.2, middle) and under the super resolution microscope(Fig.3, middle).</p>

To characterize the surface display efficiency of INP better, we added a strong promoter BBa_J23104 and RBS BBa_B0032 to the upstream of INP, and changed the YFP to sfGFP, to construct our parts BBa_K2833009(Fig.1)

<figure-class="text-center"> 

               <img src="T--BJRS_China--improvement009.png"width="50%" >
<figcaption>
Figure 1 the framework of BBa_K2833009.
</figcaption> 
           </figure>

Results

The overnight cultured bacteria expressing GFP, INP-YFP and INP-GFP respectively were firstly adjusted to OD600 = 1.0 to assure the same cell counts, then 1 mL bacteria solution of each was centrifuged to get the sediment of whole cell. From the fluorescence strength of the of whole cell(E.coli, DH5α) precipitation under UV-light we can see that the BBa_K523013 transformed cells showed the weakest fluorescence, suggesting that our new construct can give out stronger fluorescence.</p> <figure class="text-center"> <img src="T--BJRS_China--improvement1.jpg" width="50%" > <figcaption>

Figure 2 the fluorescence of whole cell(E.coli, DH5α) precipitation under UV-light A: intracellular expressing GFP bacteria; B: BBa_K523013 transformed bacteria; C:BBa_K2833009 transformed bacteria
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           </figure>

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