Difference between revisions of "Part:BBa K2336035"
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<h1>'''improvement'''</h1>
 
<h1>'''improvement'''</h1>
−
<p>  This part is an expansion of our team's work in 2015(<a href="https://parts.igem.org/Part:BBa_K1592007">BBa_K1592007</a>). Two years ago, we did it in yeast. Now we put gene into <I>E.coli</I>. It's a hard experiment and we finally make it. The gene order is also different from that in eukaryotes, yeast for instance. We consider it as a big breakthrough.</p>
+
<p>  This part is an expansion of our team's work in 2015(<li><a href="https://parts.igem.org/Part:BBa_K1592007">BBa_K1592007</a></li>). Two years ago, we did it in yeast. Now we put gene into <I>E.coli</I>. It's a hard experiment and we finally make it. The gene order is also different from that in eukaryotes, yeast for instance. We consider it as a big breakthrough.</p>
 
<p>  Next we will do experiment to test its binding ability. We hope it will have the effect as we expected.</p>
 
<p>  Next we will do experiment to test its binding ability. We hope it will have the effect as we expected.</p>
  

Revision as of 17:51, 31 October 2017


oprF-GS-Sitag-FLAG

The report part of our circuit. It can be expressed to bind silicon

Usage and Biology

oprF is motif cell wall anchor domain protein. GS stands for GS linker. Sitag-FLAG is designed to express specially protein. The genetic engineering bacteria can produce the protein on its membrane. Then the bacteria can be absorbed by silica surface. So we can capture them. Figure 1 shows the circuit of it.

Figure 1: The circuit of oprF-GS-Sitag-FLAG.

As we have said, this part is a final part. After sensing and capture finish, the bacteria will be bound onto silica surface. We can achieve our goal to get them(with lanthanum). Figure 2 shows how it works.

Figure 2: The preview of Sitag displayed on cell wall and work.

Characterization

We did some experiment to prove it works.

SDS-PAGE

After oprF-GS-Sitag-FLAG is expressed successfully,we centrifuged the bacteria liquid and separated different proteins by SDS-PAGE.

Figure 2: The preview of Sitag displayed on cell wall and work.

Figure 3 shows an obvious ~57kDa protein bands of oprF-Sitag in test lane, which cannot be found in control lane. This result proves that the bacteria could express oprF-Sitag successfully.

improvement

This part is an expansion of our team's work in 2015(

  • <a href="https://parts.igem.org/Part:BBa_K1592007">BBa_K1592007</a>
    • ). Two years ago, we did it in yeast. Now we put gene into E.coli. It's a hard experiment and we finally make it. The gene order is also different from that in eukaryotes, yeast for instance. We consider it as a big breakthrough.

    Next we will do experiment to test its binding ability. We hope it will have the effect as we expected.

    Sequence and Features


    Assembly Compatibility:
    • 10
      COMPATIBLE WITH RFC[10]
    • 12
      INCOMPATIBLE WITH RFC[12]
      Illegal NheI site found at 72
    • 21
      INCOMPATIBLE WITH RFC[21]
      Illegal BamHI site found at 639
    • 23
      COMPATIBLE WITH RFC[23]
    • 25
      INCOMPATIBLE WITH RFC[25]
      Illegal NgoMIV site found at 3
      Illegal AgeI site found at 1527
    • 1000
      COMPATIBLE WITH RFC[1000]