Difference between revisions of "Part:BBa K5526000"
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===Usage and Biology===
 
  
 
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===Functional Parameters===
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    <title>BBa_K5526000 - Azurin Documentation</title>
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    <h2>New Basic Part: BBa_K5526000 (Azurin)</h2>
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    <h3>Profile</h3>
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    <p><strong>Name:</strong> Azurin</p>
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    <p><strong>Base Pairs:</strong> 444bp</p>
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    <p><strong>Origin:</strong> <i>Pseudomonas aeruginosa</i>, synthesized.</p>
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    <p><strong>Properties:</strong> Azurin is a cupredoxin protein isolated from <i>Pseudomonas aeruginosa</i> that can enter cancer cells and induce apoptosis.</p>
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    <!-- Usage and Biology Section -->
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    <h3>Usage and Biology</h3>
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    <p>Azurin is a cupredoxin protein isolated from <i>Pseudomonas aeruginosa</i> that can enter cancer cells and induce apoptosis. It interacts with specific molecules on the surface of cancer cells, disrupting intracellular signalling pathways. Azurin is primarily in the stages of basic research and preclinical trials and has not yet become a standard clinical treatment.</p>
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        <img src="https://static.igem.wiki/teams/5526/bba-k5526000/1.png" style="width: 50%;" alt="Gene map of Azurin">
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        <figcaption>Figure 1. Gene map of Azurin</figcaption>
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    <h3>Cultivation, Purification, and SDS-PAGE</h3>
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    <p>Azurin is a gene that encodes a chemical drug, which can release substances to kill tumor cells. We applied PCR on the genes Azurin (444bp) and pUC57-plldr (3150bp). Agarose gel electrophoresis was used to check the length of our PCR production to ensure success. The result shows that the pUC57-plldr (plactate 1) has a length of 3150bp, and the Azurin has a length of 444bp.</p>
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        <img src="https://static.igem.wiki/teams/5526/bba-k5526000/2.jpg" style="width: 50%;" alt="PCR Production Identification">
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        <figcaption>Figure 2. The identification of PCR production by agarose gel electrophoresis. Left: The graph shows that plactate 1 has a length of 3150bp. Right: The graph shows p1-Azurin has a length of 444bp.</figcaption>
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    <p>After the optimal protein expression conditions were obtained, the constructed strains were expanded for culture. 5 mM lactic acid was used to induce the actual expression level of tumor drugs in EcN1917. The experimental results are shown in Figure 3. The size of the Azurin protein is about 19kDa. A large amount of target protein indicates that the Azurin protein is successfully expressed. In the future, we will further study the expression and tumor inhibition effect of the constructed probiotics in the tumor environment.</p>
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        <img src="https://static.igem.wiki/teams/5526/bba-k5526000/3.jpg" style="width: 50%;" alt="Azurin protein expression by SDS-PAGE">
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        <figcaption>Figure 3. Detection of Azurin protein expression by SDS-PAGE</figcaption>
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Revision as of 14:01, 28 September 2024

Azurin

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]


BBa_K5526000 - Azurin Documentation

New Basic Part: BBa_K5526000 (Azurin)

Profile

Name: Azurin

Base Pairs: 444bp

Origin: Pseudomonas aeruginosa, synthesized.

Properties: Azurin is a cupredoxin protein isolated from Pseudomonas aeruginosa that can enter cancer cells and induce apoptosis.

Usage and Biology

Azurin is a cupredoxin protein isolated from Pseudomonas aeruginosa that can enter cancer cells and induce apoptosis. It interacts with specific molecules on the surface of cancer cells, disrupting intracellular signalling pathways. Azurin is primarily in the stages of basic research and preclinical trials and has not yet become a standard clinical treatment.

Gene map of Azurin
Figure 1. Gene map of Azurin

Cultivation, Purification, and SDS-PAGE

Azurin is a gene that encodes a chemical drug, which can release substances to kill tumor cells. We applied PCR on the genes Azurin (444bp) and pUC57-plldr (3150bp). Agarose gel electrophoresis was used to check the length of our PCR production to ensure success. The result shows that the pUC57-plldr (plactate 1) has a length of 3150bp, and the Azurin has a length of 444bp.

PCR Production Identification
Figure 2. The identification of PCR production by agarose gel electrophoresis. Left: The graph shows that plactate 1 has a length of 3150bp. Right: The graph shows p1-Azurin has a length of 444bp.

After the optimal protein expression conditions were obtained, the constructed strains were expanded for culture. 5 mM lactic acid was used to induce the actual expression level of tumor drugs in EcN1917. The experimental results are shown in Figure 3. The size of the Azurin protein is about 19kDa. A large amount of target protein indicates that the Azurin protein is successfully expressed. In the future, we will further study the expression and tumor inhibition effect of the constructed probiotics in the tumor environment.

Azurin protein expression by SDS-PAGE
Figure 3. Detection of Azurin protein expression by SDS-PAGE