Difference between revisions of "Part:BBa K4606010"
 
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==Description==
 
==Description==
 
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     <p>For detailed design information, you can look up the registry of BBa_K4606007, BBa_K4606001 and BBa_K4606005.</p>
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     <p>For detailed design information, you can look up the registry of <a href="https://parts.igem.org/Part:BBa_K4606007">BBa_K4606007</a>, <a href="https://parts.igem.org/Part:BBa_K4606001">BBa_K4606001</a> and ,<a href="https://parts.igem.org/Part:BBa_K4606005">BBa_K4606005.</a> </p>
 
     <p>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.</p>
 
     <p>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.</p>
 
     <p>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.</p>
 
     <p>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.</p>
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         <p>Figure 1. Expression map of the composite plasmids of miR-3074-5p.</p>
 
         <p>Figure 1. Expression map of the composite plasmids of miR-3074-5p.</p>
 
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    <div style="margin:auto; width: 400px">   
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        <img src="https://static.igem.wiki/teams/4606/wiki/part-1-zsgreen.jpg" width="400px">
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    <div style="margin:auto; width:400px;">
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        <p>Figure 2. After the plasmid was transfected into the cells, the fluorescent protein was expressed and emitted green fluorescence.</p>
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    <div style="margin:auto; width: 250px">   
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        <img src="https://static.igem.wiki/teams/4606/wiki/part-5-figure1.jpg" width="250px">
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    </div>
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    <div style="margin:auto; width:400px;">
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        <p>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.</p>
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    </div>
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==Design notes==
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    <p>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.</p>
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==Source==
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    <p>Plasmids were obtained from Escherichia coli.</p>
 
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Latest revision as of 06:58, 23 September 2023


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


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]