Difference between revisions of "Part:BBa K4606007"

 
 
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<partinfo>BBa_K4606007 short</partinfo>
 
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==description==
  
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    <p>In this part, we designed a plasmid for fusion protein recombination. With this device, we can insert the coding sequence of a novel protein before Poly A. When transfected into E. coli or other recipient cells, it amplifies and expresses the corresponding gene product.</p>
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    <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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    <div style="margin:auto; width: 400px">   
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        <img src="https://static.igem.wiki/teams/4606/wiki/part-7-linear-vector-plasmid.jpg" width="400px">
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    </div>
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    <div style="margin:auto; width:400px;">
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        <p>Figure 1. Restriction digest verification of vector plasmid by PCR.
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            <br>1#: DNA marker. 2#: Control vector plasmid. 3-5#: Enzyme digestion product of the vector plasmid.
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        </p>
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    </div>
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==Usage==
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    <p>To use this device, all you need to do is cut the plasmid at Xba1 and insert your target fragment into this position. We have proved its feasibility by inserting our destination fragment (Firefly Luciferase) to the plasmid. The recombinants show resistance to Ampicillin. </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-vector-plasmid-map.jpg" width="400px">
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    </div>
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    <div style="margin:auto; width:400px;">
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        <p>Figure 2. Expression map of the vector plasmids.</p>
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    </div>
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===Usage and Biology===
 
  
 
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Latest revision as of 06:37, 23 September 2023


Vector-AmpR

description

In this part, we designed a plasmid for fusion protein recombination. With this device, we can insert the coding sequence of a novel protein before Poly A. When transfected into E. coli or other recipient cells, it amplifies and expresses the corresponding gene product.

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.

Figure 1. Restriction digest verification of vector plasmid by PCR.
1#: DNA marker. 2#: Control vector plasmid. 3-5#: Enzyme digestion product of the vector plasmid.

Usage

To use this device, all you need to do is cut the plasmid at Xba1 and insert your target fragment into this position. We have proved its feasibility by inserting our destination fragment (Firefly Luciferase) to the plasmid. The recombinants show resistance to Ampicillin.

Figure 2. Expression map of the vector plasmids.

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]