Difference between revisions of "Part:BBa K5302026"

 
 
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This work is derived from pBBR1MCS-OmpA-mCherry and pUC19-8IJZ, and it has undergone codon optimization. This composite part combines OmpA(21.4kda) and 8IJZ, we succeeded in transferring this plasmid into Escherichia coli Nissle 1917 and let it express 8IJZ. The plasmid uses lac promotor and has kanamycin resistence.
 
This work is derived from pBBR1MCS-OmpA-mCherry and pUC19-8IJZ, and it has undergone codon optimization. This composite part combines OmpA(21.4kda) and 8IJZ, we succeeded in transferring this plasmid into Escherichia coli Nissle 1917 and let it express 8IJZ. The plasmid uses lac promotor and has kanamycin resistence.
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    <img src="https://static.igem.wiki/teams/5302/images/part-registry-8ijz-2.jpg"
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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> Colony PCR results of pBBR-INP-8IJZ
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8IJZ is an anti-VEGF nanobody, and it has 2 chains.These two chains all show great affinity with VEGF, we plan to use pBBR1MCS-2 plasmid as a backbone and transfer 8IJZ into Escherichia coli Nissle 1917, and finally succeeded in expressing 8IJZ.
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    <img src="https://static.igem.wiki/teams/5302/images/part-registry-8ijz-1.jpg"
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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> structure of 8IJZ， from PDB
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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 02:00, 2 October 2024


pBBR-INP-8IJZ

This work is derived from pBBR1MCS-OmpA-mCherry and pUC19-8IJZ, and it has undergone codon optimization. This composite part combines OmpA(21.4kda) and 8IJZ, we succeeded in transferring this plasmid into Escherichia coli Nissle 1917 and let it express 8IJZ. The plasmid uses lac promotor and has kanamycin resistence.

Jamboree Program
Figure 1. Colony PCR results of pBBR-INP-8IJZ

8IJZ is an anti-VEGF nanobody, and it has 2 chains.These two chains all show great affinity with VEGF, we plan to use pBBR1MCS-2 plasmid as a backbone and transfer 8IJZ into Escherichia coli Nissle 1917, and finally succeeded in expressing 8IJZ.

Jamboree Program
Figure 2. structure of 8IJZ， from PDB

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 4659
    Illegal XbaI site found at 3189
    Illegal PstI site found at 2016
    Illegal PstI site found at 3177
    Illegal PstI site found at 4427
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 4659
    Illegal PstI site found at 2016
    Illegal PstI site found at 3177
    Illegal PstI site found at 4427
    Illegal NotI site found at 1057
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 4659
    Illegal BglII site found at 1803
    Illegal BamHI site found at 3195
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 4659
    Illegal XbaI site found at 3189
    Illegal PstI site found at 2016
    Illegal PstI site found at 3177
    Illegal PstI site found at 4427
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 4659
    Illegal XbaI site found at 3189
    Illegal PstI site found at 2016
    Illegal PstI site found at 3177
    Illegal PstI site found at 4427
    Illegal NgoMIV site found at 2467
    Illegal NgoMIV site found at 2750
    Illegal NgoMIV site found at 3616
    Illegal NgoMIV site found at 5324
    Illegal AgeI site found at 5164
  • 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