Difference between revisions of "Part:BBa K5302020"

 
 
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Just like miniZ, Z3C is also derived from three helix 58-residue Z-domain of staphylococcal protein A, but it has three helix. Z3C also shows great affinity with VEGF(KD=41 nM).  
 
Just like miniZ, Z3C is also derived from three helix 58-residue Z-domain of staphylococcal protein A, but it has three helix. Z3C also shows great affinity with VEGF(KD=41 nM).  
  
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We used pBBR plasmid as a backbone and transfered Z3C into Escherichia coli Nissle 1917, and finally succeeded in expressing Z3C.
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We used pBBR1MCS-2 plasmid as a backbone and transfered Z3C into Escherichia coli Nissle 1917, and finally succeeded in expressing Z3C.
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<div style="text-align:center;">
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    <img src="https://static.igem.wiki/teams/5302/images/part-registry-poz3c-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> Colony PCR results of pBBR-OmpA-Z3C
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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-poz3c-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> SDS-PAGE analysis of pBBR1MCS-OmpA-Z3C expression in Escherichia coli Nissle 1917 （1）
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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-poz3c-3.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 3. </b> SDS-PAGE analysis of pBBR1MCS-OmpA-Z3C expression in Escherichia coli Nissle 1917（2）
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        </caption>
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    </div>
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Z3C is derived from three helix 58-residue Z-domain of staphylococcal protein A, but it has three helix. Z3C also shows great affinity with VEGF(KD=41 nM). We used pBBR plasmid as a backbone and transfered Z3C into Escherichia coli Nissle 1917, and finally succeeded in expressing Z3C.
 +
 
 +
<html>
 +
<div style="text-align:center;">
 +
    <img src="https://static.igem.wiki/teams/5302/images/part-registry-z3c-1.png"
 +
        width="60%" style="display:block; margin:auto;" alt="Jamboree Program" >
 +
    <div style="text-align:center;">
 +
        <caption>
 +
            <b>Figure 4. </b> sequence of Z3C and its KD with VEGF
 +
        </caption>
 +
    </div>
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</div>
 +
</html>
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 +
<html>
 +
<div style="text-align:center;">
 +
    <img src="https://static.igem.wiki/teams/5302/images/part-registry-z3c-2.png"
 +
        width="60%" style="display:block; margin:auto;" alt="Jamboree Program" >
 +
    <div style="text-align:center;">
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        <caption>
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            <b>Figure 5. </b> Cartoon representation of the crystal structure of the mini-Z highlighting randomized residues shown as sticks and coloring the helices
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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 01:44, 2 October 2024


pBBR-OmpA-Z3C

Just like miniZ, Z3C is also derived from three helix 58-residue Z-domain of staphylococcal protein A, but it has three helix. Z3C also shows great affinity with VEGF(KD=41 nM).

We used pBBR1MCS-2 plasmid as a backbone and transfered Z3C into Escherichia coli Nissle 1917, and finally succeeded in expressing Z3C.

Jamboree Program
Figure 1. Colony PCR results of pBBR-OmpA-Z3C

Jamboree Program
Figure 2. SDS-PAGE analysis of pBBR1MCS-OmpA-Z3C expression in Escherichia coli Nissle 1917 （1）

Jamboree Program
Figure 3. SDS-PAGE analysis of pBBR1MCS-OmpA-Z3C expression in Escherichia coli Nissle 1917（2）

Z3C is derived from three helix 58-residue Z-domain of staphylococcal protein A, but it has three helix. Z3C also shows great affinity with VEGF(KD=41 nM). We used pBBR plasmid as a backbone and transfered Z3C into Escherichia coli Nissle 1917, and finally succeeded in expressing Z3C.

Jamboree Program
Figure 4. sequence of Z3C and its KD with VEGF

Jamboree Program
Figure 5. Cartoon representation of the crystal structure of the mini-Z highlighting randomized residues shown as sticks and coloring the helices

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 4100
    Illegal XbaI site found at 3140
    Illegal PstI site found at 1967
    Illegal PstI site found at 3128
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 4100
    Illegal PstI site found at 1967
    Illegal PstI site found at 3128
    Illegal NotI site found at 1008
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 4100
    Illegal BglII site found at 1754
    Illegal BamHI site found at 3146
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 4100
    Illegal XbaI site found at 3140
    Illegal PstI site found at 1967
    Illegal PstI site found at 3128
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 4100
    Illegal XbaI site found at 3140
    Illegal PstI site found at 1967
    Illegal PstI site found at 3128
    Illegal NgoMIV site found at 2418
    Illegal NgoMIV site found at 2701
    Illegal NgoMIV site found at 4758
    Illegal AgeI site found at 3789
    Illegal AgeI site found at 4598
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1330
    Illegal SapI.rc site found at 2267
    Illegal SapI.rc site found at 2477