Difference between revisions of "Part:BBa K5074001"
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According to the aptamer sequence of Shigella dysenteriae and the construction rules of toehold switch, we designed the aptamer toehold switch with amilCP reporter gene, which was aimed to detect Shigella dysenteriae.  
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In order to make the detection of pathogenic bacteria more intuitive and visible, we used some chromoproteins as the reporters. One of which is AmilCP(K592009) which shows purple chromoprotein used to construct BBa_K5074001.
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According to the aptamer sequence of Shigella dysenteriae and the construction rules of toehold switch, we designed the aptamer toehold switch with amilCP reporter gene, which was aimed to detect Shigella dysenteriae in refrigerator.
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This part comprises Shigella dysenteriae RNA aptamer, RBS(B0034), Linker and reporter gene amilCP(K592009). It’s sequence was analyzed using Snapgene Viewer for restriction enzyme cleaving sites to congstruct the plasmid pET-28a-Shig-amilCP using the proper enzymes. The analysis result with Snapgene Viewer was shown in Fig.1.  
  
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It comprises Shigella dysenteriae RNA aptamer, RBS, Linker and reporter gene amilCP. It’s sequence was analyzed with Snapgene Viewer for restriction enzyme cleaving sites to congstruct the plasmid using the proper enzymes. The analysis result of Snapgene was shown in Fig.1.
 
  
 
<p align="center">https://static.igem.wiki/teams/5074/part/part-k5074001-fig-1-2.jpg</p >           
 
<p align="center">https://static.igem.wiki/teams/5074/part/part-k5074001-fig-1-2.jpg</p >           
  
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Fig.1 The map of BBa_K5074001 analized with SnapGene Viewer, showing the restriction enzyme information (no EcoRI and PstI sites).
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Fig.1 The endonuclease map of BBa_K5074001 analized using SnapGene Viewer, showing the restriction enzyme information (no EcoRI and Hind III cleaving sites).

Revision as of 06:25, 30 September 2024


AmilCP generator controlled by Shigella RNA aptamer toehold switch

It comprises Shigella dysenteriae RNA aptamer, RBS, Linker and reporter gene amilCP. At the presence of Shigella dysenteriae, it binds to its RNA aptamer, opening the toehold switch to trigger the expression of amilCP, which is easily deteced. It is used to detect the bacteria Shigella in refrigerator.

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]


In order to make the detection of pathogenic bacteria more intuitive and visible, we used some chromoproteins as the reporters. One of which is AmilCP(K592009) which shows purple chromoprotein used to construct BBa_K5074001.

According to the aptamer sequence of Shigella dysenteriae and the construction rules of toehold switch, we designed the aptamer toehold switch with amilCP reporter gene, which was aimed to detect Shigella dysenteriae in refrigerator.

This part comprises Shigella dysenteriae RNA aptamer, RBS(B0034), Linker and reporter gene amilCP(K592009). It’s sequence was analyzed using Snapgene Viewer for restriction enzyme cleaving sites to congstruct the plasmid pET-28a-Shig-amilCP using the proper enzymes. The analysis result with Snapgene Viewer was shown in Fig.1.


part-k5074001-fig-1-2.jpg

Fig.1 The endonuclease map of BBa_K5074001 analized using SnapGene Viewer, showing the restriction enzyme information (no EcoRI and Hind III cleaving sites).