Difference between revisions of "Part:BBa K4134077"

 
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<partinfo>BBa_K4134077 short</partinfo>
 
<partinfo>BBa_K4134077 short</partinfo>
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==Description==
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Almost the same as our another parts BBa_K4134066. The only change is that we replace Atox1 with AgBP2 as silver binding protein. But as devices for fusion protein recombination, both of them function the same.  
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[[Image:Nanjing-China-recombination-device-AgBP2.jpeg|400px|thumb|right|Homologous recombination device design]]
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In detail, we designed a device for fusion protein recombination. With this device, you can display any small protein to the C-terminus of BpfA, a large membrane protein of S. oneidensis.  
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<html><p>
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Considering the addition of KanR, you can easily confirm whether homologous recombination succeeds or not, since only the recombinants can grow into colonies on the medium supplemented with kanamycin.
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Almost the same as our another part <a href="https://parts.igem.org/Part:BBa_K4134066"><strong>BBa_K4134066</strong></a>. The only change is that we replace Atox1 with AgBP2 as silver binding protein. But as devices for fusion protein recombination, both of them function the same. </p>
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To use this device, all you need to do is to replace AgBP2 with your destination fragment. We have proved its feasibility by fusing our destination fragment (AgBP2) to the C-terminus of BpfA. The recombinants show resistance to kanamycin and enrich silver ions on the cell membrane.  
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<p>
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Moreover, there is a flexible linker between BpfA and your destination fragment. Linker is an amino acid chain that acts as a link between two fusion proteins and is flexible enough to allow the proteins on both sides to perform their independent functions.   
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In detail, we designed a device for fusion protein recombination. With this device, you can display any small protein to the C-terminus of BpfA, a large membrane protein of S. oneidensis. </p>
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In detail, BpfA stands for the biofilm-promoting protein A, a large surface protein. The 1000bp-long fragment of BpfA and the 1000bp-long fragment of AggC, the downstream genes of BpfA, are attached to our vector for homologous recombination so as to fuse the destination fragment to the C-terminus of BpfA. This design enables the destination gene to be displayed on the surface of S. oneidensis. Considering the large size of BpfA, this fusion expression does nearly no harm to the normal physiological state of S. oneidensis and we have approved it through a biofilm growth test and bacterial density measurement.  
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<p>
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The loxP site-specific recombination site regulates the insertion of the destination fragment at the specific location bpfA, ensuring the fusion of it to bpfA through homologous recombination.
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Considering the addition of KanR, you can easily confirm whether homologous recombination succeeds or not, since only the recombinants can grow into colonies on the medium supplemented with kanamycin.</p>
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+
<p>
 +
To use this device, all you need to do is to replace AgBP2 with your destination fragment. We have proved its feasibility by fusing our destination fragment (AgBP2) to the C-terminus of BpfA. The recombinants show resistance to kanamycin and enrich silver ions on the cell membrane. </p>
 +
<p>
 +
Moreover, there is a flexible linker between your destination fragment and KanR. Linker is an amino acid chain that acts as a link between two fusion proteins and is flexible enough to allow the proteins on both sides to perform their independent functions.  </p>
 +
<p>
 +
In detail, BpfA stands for the biofilm-promoting protein A, a large surface protein. The 1000bp-long fragment of BpfA and the 1000bp-long fragment of AggC, the downstream genes of BpfA, are attached to our vector for homologous recombination so as to fuse the destination fragment to the C-terminus of BpfA. This design enables the destination gene to be displayed on the surface of S. oneidensis. Considering the large size of BpfA, this fusion expression does nearly no harm to the normal physiological state of S. oneidensis and we have approved it through a biofilm growth test and bacterial density measurement. </p>
 +
<p>
 +
The loxP site-specific recombination site regulates the insertion of the destination fragment at the specific location bpfA, ensuring the fusion of it to bpfA through homologous recombination.</p>
 +
</html>
  
 
<!-- Add more about the biology of this part here
 
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Latest revision as of 14:50, 13 October 2022


BpfA-AgBP2-Linker-KanR-loxP-AggC

Description

Homologous recombination device design

Almost the same as our another part BBa_K4134066. The only change is that we replace Atox1 with AgBP2 as silver binding protein. But as devices for fusion protein recombination, both of them function the same.

In detail, we designed a device for fusion protein recombination. With this device, you can display any small protein to the C-terminus of BpfA, a large membrane protein of S. oneidensis.

Considering the addition of KanR, you can easily confirm whether homologous recombination succeeds or not, since only the recombinants can grow into colonies on the medium supplemented with kanamycin.

To use this device, all you need to do is to replace AgBP2 with your destination fragment. We have proved its feasibility by fusing our destination fragment (AgBP2) to the C-terminus of BpfA. The recombinants show resistance to kanamycin and enrich silver ions on the cell membrane.

Moreover, there is a flexible linker between your destination fragment and KanR. Linker is an amino acid chain that acts as a link between two fusion proteins and is flexible enough to allow the proteins on both sides to perform their independent functions.

In detail, BpfA stands for the biofilm-promoting protein A, a large surface protein. The 1000bp-long fragment of BpfA and the 1000bp-long fragment of AggC, the downstream genes of BpfA, are attached to our vector for homologous recombination so as to fuse the destination fragment to the C-terminus of BpfA. This design enables the destination gene to be displayed on the surface of S. oneidensis. Considering the large size of BpfA, this fusion expression does nearly no harm to the normal physiological state of S. oneidensis and we have approved it through a biofilm growth test and bacterial density measurement.

The loxP site-specific recombination site regulates the insertion of the destination fragment at the specific location bpfA, ensuring the fusion of it to bpfA through homologous recombination.

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]