Part:BBa_K5321018
βhCG_PPV cleavage site_GST
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 691
Illegal NgoMIV site found at 874
Illegal NgoMIV site found at 918 - 1000INCOMPATIBLE 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.
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