Difference between revisions of "Part:BBa K5302030"

 
 
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This work is derived from pBBR1MCS-INP-mCherry and PUC19-ZVEGF-V114-V107-peptide, and it has undergone codon optimization. This composite part combines INP(about 30kda) and V114(approximately 2.3kda), we succeeded in transferring this plasmid into Escherichia coli Nissle 1917 and let it express V114. The plasmid uses lac promotor and has kanamycin resistence.
 
This work is derived from pBBR1MCS-INP-mCherry and PUC19-ZVEGF-V114-V107-peptide, and it has undergone codon optimization. This composite part combines INP(about 30kda) and V114(approximately 2.3kda), we succeeded in transferring this plasmid into Escherichia coli Nissle 1917 and let it express V114. The plasmid uses lac promotor and has kanamycin resistence.
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<hr>
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V114 has been identified as a small potent VEGF-binding peptide, which is composed of 8 amino acids--- that is VEPNc[CDIHVnLWEWEC]FERL, and there is a disulfide bond between N and F, making the stucture more stable. This peptide has a similiar structure and VEGF-binding site as V107, and it shows great affinity with VEGF(Kd=0.11 μM). We used pBBR1MCS-2 plasmid as a backbone and transfered V114 into Escherichia coli Nissle 1917, and finally succeeded in expressing V114.
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<div style="text-align:center;">
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    <img src="https://static.igem.wiki/teams/5302/images/part-registry-v114-1.png"
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        width="60%" style="display:block; margin:auto;" alt="Jamboree Program" >
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    <div style="text-align:center;">
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        <caption>
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            <b>Figure 1. </b> This picture shows its VEGF-binding site and its affinity with VEGF
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        </caption>
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    </div>
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</div>
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</html>
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<html>
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<div style="text-align:center;">
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    <img src="https://static.igem.wiki/teams/5302/images/part-registry-v114-2.png"
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        width="60%" style="display:block; margin:auto;" alt="Jamboree Program" >
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    <div style="text-align:center;">
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        <caption>
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            <b>Figure 2. </b> This picture shows the dependence of the amount of V114, expressed in normalized fluorescence values, on the peptide concentration
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        </caption>
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    </div>
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</div>
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</html>
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 02:06, 2 October 2024


pBBR-INP-V114

This work is derived from pBBR1MCS-INP-mCherry and PUC19-ZVEGF-V114-V107-peptide, and it has undergone codon optimization. This composite part combines INP(about 30kda) and V114(approximately 2.3kda), we succeeded in transferring this plasmid into Escherichia coli Nissle 1917 and let it express V114. The plasmid uses lac promotor and has kanamycin resistence.

V114 has been identified as a small potent VEGF-binding peptide, which is composed of 8 amino acids--- that is VEPNc[CDIHVnLWEWEC]FERL, and there is a disulfide bond between N and F, making the stucture more stable. This peptide has a similiar structure and VEGF-binding site as V107, and it shows great affinity with VEGF(Kd=0.11 μM). We used pBBR1MCS-2 plasmid as a backbone and transfered V114 into Escherichia coli Nissle 1917, and finally succeeded in expressing V114.

Jamboree Program
Figure 1. This picture shows its VEGF-binding site and its affinity with VEGF

Jamboree Program
Figure 2. This picture shows the dependence of the amount of V114, expressed in normalized fluorescence values, on the peptide concentration

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 4338
    Illegal XbaI site found at 3189
    Illegal PstI site found at 2016
    Illegal PstI site found at 3177
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 4338
    Illegal PstI site found at 2016
    Illegal PstI site found at 3177
    Illegal NotI site found at 1057
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 4338
    Illegal BglII site found at 1803
    Illegal BamHI site found at 3195
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 4338
    Illegal XbaI site found at 3189
    Illegal PstI site found at 2016
    Illegal PstI site found at 3177
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 4338
    Illegal XbaI site found at 3189
    Illegal PstI site found at 2016
    Illegal PstI site found at 3177
    Illegal NgoMIV site found at 2467
    Illegal NgoMIV site found at 2750
    Illegal NgoMIV site found at 3616
    Illegal NgoMIV site found at 5003
    Illegal AgeI site found at 4843
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1379
    Illegal SapI.rc site found at 2316
    Illegal SapI.rc site found at 2526