Difference between revisions of "Part:BBa K5241005"

 
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Based on Pseudomonas aeruginosa PAO1 [gene=hppD] [protein=4-hydroxyphenylpyruvate dioxygenase], a Pahdd-6xH gene with a 6xH tag is synthesized in pET-21b(+), with the anticipated protein localization within the bacterial cytoplasm.
 
Based on Pseudomonas aeruginosa PAO1 [gene=hppD] [protein=4-hydroxyphenylpyruvate dioxygenase], a Pahdd-6xH gene with a 6xH tag is synthesized in pET-21b(+), with the anticipated protein localization within the bacterial cytoplasm.
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Gene Circuit Diagram:
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<figure><img style="width: 65%; padding:28px;"src="https://static.igem.wiki/teams/5241/wiki/parts-static/5/bba-k5241005-img01.png"></figure>
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</html>
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Result：We successfully expressed the hppD gene using IPTG induction, and the 2×BL21(DE3) strain performed better than the 2×BL21(DE3) pLysS; the expression of melanin was more effective than that of the PkhppD gene.
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<html>
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<figure>
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<img style="width: 48%; padding:28px 2px;"src="https://static.igem.wiki/teams/5241/wiki/parts-static/5/bba-k5241005-img02.png">
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<img style="width: 48%; padding:28px 2px;"src="https://static.igem.wiki/teams/5241/wiki/parts-static/5/bba-k5241005-img03.png">
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</figure>
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</html>
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Reference documentation
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[1]ChiaraScanferla EnricoCaruso Viviana TeresaOrlandi FabrizioBolognese. Bacterial melanin production by heterologous expression of 4‑hydroxyphenylpyruvate dioxygenase from Pseudomonas aeruginosa[J]. International Journal of Biological Macromolecules, 2019, 133: 1072-1080.
  
 
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Revision as of 07:50, 27 September 2024


Pahdd-6xH in pET-21b(+)

Based on Pseudomonas aeruginosa PAO1 [gene=hppD] [protein=4-hydroxyphenylpyruvate dioxygenase], a Pahdd-6xH gene with a 6xH tag is synthesized in pET-21b(+), with the anticipated protein localization within the bacterial cytoplasm.

Gene Circuit Diagram:

Result：We successfully expressed the hppD gene using IPTG induction, and the 2×BL21(DE3) strain performed better than the 2×BL21(DE3) pLysS; the expression of melanin was more effective than that of the PkhppD gene.

Reference documentation

[1]ChiaraScanferla EnricoCaruso Viviana TeresaOrlandi FabrizioBolognese. Bacterial melanin production by heterologous expression of 4‑hydroxyphenylpyruvate dioxygenase from Pseudomonas aeruginosa[J]. International Journal of Biological Macromolecules, 2019, 133: 1072-1080.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 2950
    Illegal EcoRI site found at 3121
    Illegal EcoRI site found at 3595
    Illegal PstI site found at 3826
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 2950
    Illegal EcoRI site found at 3121
    Illegal EcoRI site found at 3595
    Illegal PstI site found at 3826
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 2950
    Illegal EcoRI site found at 3121
    Illegal EcoRI site found at 3595
    Illegal BglII site found at 587
    Illegal BglII site found at 2316
    Illegal BglII site found at 3830
    Illegal BamHI site found at 562
    Illegal BamHI site found at 1630
    Illegal XhoI site found at 676
    Illegal XhoI site found at 1624
    Illegal XhoI site found at 3763
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 2950
    Illegal EcoRI site found at 3121
    Illegal EcoRI site found at 3595
    Illegal PstI site found at 3826
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 2950
    Illegal EcoRI site found at 3121
    Illegal EcoRI site found at 3595
    Illegal PstI site found at 3826
    Illegal AgeI site found at 524
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 75
    Illegal BsaI site found at 1388
    Illegal BsaI.rc site found at 2149
    Illegal BsaI.rc site found at 2617