Part:BBa_K4606010
Vector-ZsGreen-miR-3074-5p-AmpR
Description
For detailed design information, you can look up the registry of BBa_K4606007, BBa_K4606001 and ,BBa_K4606005.
Here, we loaded the genes encoding miR-3074-5p into plasmid vectors. In this way we were able to over-express the miRNA in the target cells.
Due to the addition of the AmpR gene to the plasmid, we could easily confirm whether the recombination was successful or not, as only the recombinants can grow into colonies on the medium supplemented with Ampicillin.
ZsGreen protein is spontaneously expressed and emits green fluorescence after successful transfection, so we can observe the transfection efficiency of the vector.
Figure 1. Expression map of the composite plasmids of miR-3074-5p.
Figure 2. After the plasmid was transfected into the cells, the fluorescent protein was expressed and emitted green fluorescence.
Figure 3. The relative miR-3074-5p level in chondrocytes transfected with overexpression (OE) plasmid was significantly higher than that of negative control (NC) plasmid.
Design notes
This device is used to transfer the coding gene of miR-3074-5p into chondrocytes. Due to the addition of the AmpR gene to the plasmid, we could easily confirm whether the recombination was successful or not, as only the recombinants can grow into colonies on the medium supplemented with Ampicillin. ZsGreen protein is spontaneously expressed and emits green fluorescence after successful transfection, so we can observe the transfection efficiency of the vector.
Source
Plasmids were obtained from Escherichia coli.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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