Difference between revisions of "Part:BBa K5321018"

 
 
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===Sequence and Features===
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===Usage and Biology===
  
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βhCG_PPV cleavage site_GST is the substrate of PPVp in output module, and it is immobilized on a GST affinity chromatography column. Then, when our sample flows through the column, if the split-PPVp is active in the sample, cleavage happens. hCG flows out into the hCG colloidal test paper. The test paper will show a positive result. If there isn't any active PPVp in the sample, then the hCG colloidal test paper will show a negative result.
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&#946;hCG_PPV cleavage site_GST is the substrate of PPVp in output module, and it is immobilized on a GST affinity chromatography column. Then, when our sample flows through the column, if the split-PPVp is active in the sample, cleavage happens. hCG flows out into the hCG colloidal test paper. The test paper will show a positive result. If there isn't any active PPVp in the sample, then the hCG colloidal test paper will show a negative result.
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===Usage and Biology===
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<span class='h3bb'>Sequence and Features</span>
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'''Figure 1 | Concept diagram of realizing output with hCG colloidal gold test paper.'''<br>
  
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===Characterization===
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====Protein Expression====
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The plasmid containing β-hCG-GST was transformed into BL21(DE3) and protein expression was induced. We performed SDS-PAGE analysis of β-hCG, and verified its expression (Fig 3). Then, we tested whether the protein(which was expressed in the supernatant) can trigger the output signal, i.e. the positive band on the colloidal gold test paper. Unfortunatly, there’s no positive band shown.
  
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===Functional Parameters===
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'''Figure 2 | SDS-PAGE analysis of β-hCG-GST.''' The black arrow indicates the position of β-hCG-GST.<br>

Latest revision as of 03:23, 27 September 2024

βhCG_PPV cleavage site_GST

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
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 691
    Illegal NgoMIV site found at 874
    Illegal NgoMIV site found at 918
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1008
    Illegal SapI.rc site found at 85

Usage and Biology

βhCG_PPV cleavage site_GST is the substrate of PPVp in output module, and it is immobilized on a GST affinity chromatography column. Then, when our sample flows through the column, if the split-PPVp is active in the sample, cleavage happens. hCG flows out into the hCG colloidal test paper. The test paper will show a positive result. If there isn't any active PPVp in the sample, then the hCG colloidal test paper will show a negative result.


Figure 1 | Concept diagram of realizing output with hCG colloidal gold test paper.

Characterization

Protein Expression

The plasmid containing β-hCG-GST was transformed into BL21(DE3) and protein expression was induced. We performed SDS-PAGE analysis of β-hCG, and verified its expression (Fig 3). Then, we tested whether the protein(which was expressed in the supernatant) can trigger the output signal, i.e. the positive band on the colloidal gold test paper. Unfortunatly, there’s no positive band shown.


Figure 2 | SDS-PAGE analysis of β-hCG-GST. The black arrow indicates the position of β-hCG-GST.