Difference between revisions of "Part:BBa K5302027"

 
 
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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 ZVEGF(approximately 6kda), we succeeded in transferring this plasmid into Escherichia coli Nissle 1917 and let it express ZVEGF. 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 ZVEGF(approximately 6kda), we succeeded in transferring this plasmid into Escherichia coli Nissle 1917 and let it express ZVEGF. The plasmid uses lac promotor and has kanamycin resistence.
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ZVEGF is a 59-residue three-helix peptide that was developed by Fedorova et al. by randomizing 9 residues on helices 1 and 2 of the Z-domain scaffold via phage display, followed by selection for binding to the VEGF8-109 dimer. A co-crystal structure of ZVEGF with VEGF8-109 shows that the engineered Z-domain adopts the expected three-helix bundle tertiary structure and engages the receptor-recognition sites on the VEGF dimer through a surface formed by helices 1 and 2 of ZVEGF. It shows great affinity with VEGF(Ki=0.41 μM).We used pBBR1MCS-2 plasmid as a backbone and transfered ZVEGF into Escherichia coli Nissle 1917, and finally succeeded in expressing ZVEGF.
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<div style="text-align:center;">
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    <img src="https://static.igem.wiki/teams/5302/images/part-registry-v107-5.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> ZVEGF sequence
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        </caption>
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    </div>
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<div style="text-align:center;">
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    <img src="https://static.igem.wiki/teams/5302/images/part-registry-v107-6.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> VEGF-binding site of ZVEGF
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        </caption>
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    </div>
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 03:04, 2 October 2024


pBBR-INP-ZVEGF

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 ZVEGF(approximately 6kda), we succeeded in transferring this plasmid into Escherichia coli Nissle 1917 and let it express ZVEGF. The plasmid uses lac promotor and has kanamycin resistence.

ZVEGF is a 59-residue three-helix peptide that was developed by Fedorova et al. by randomizing 9 residues on helices 1 and 2 of the Z-domain scaffold via phage display, followed by selection for binding to the VEGF8-109 dimer. A co-crystal structure of ZVEGF with VEGF8-109 shows that the engineered Z-domain adopts the expected three-helix bundle tertiary structure and engages the receptor-recognition sites on the VEGF dimer through a surface formed by helices 1 and 2 of ZVEGF. It shows great affinity with VEGF(Ki=0.41 μM).We used pBBR1MCS-2 plasmid as a backbone and transfered ZVEGF into Escherichia coli Nissle 1917, and finally succeeded in expressing ZVEGF.

Jamboree Program
Figure 1. ZVEGF sequence

Jamboree Program
Figure 2. VEGF-binding site of ZVEGF

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 4458
    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 4458
    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 4458
    Illegal BglII site found at 1803
    Illegal BamHI site found at 3195
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 4458
    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 4458
    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 5123
    Illegal AgeI site found at 4963
  • 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