Difference between revisions of "Part:BBa K3598011"
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The circuit we transformed into Pichia Pastoris to produce fp151 protein.
 
The circuit we transformed into Pichia Pastoris to produce fp151 protein.
  
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===Usage and Biology===
 
===Usage and Biology===
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[[File:T--BEIJING 4ELEVEN--AOX151_1.png|600px|thumb|center|Figure 1. Part demonstration]]
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Our AOX1-fp1-mfp5-fp1 is a composite part consisting of an AOX1 Promoter, an fp1-mfp5-fp1 recombinant protein sequence, and an AOX1 Terminator. It regulates the expression of our recombinant adhesive protein. We inserted the sequence of the system onto vector pPIC9K and transferred the resulting plasmid into Pichia pastoris inoculated on BMMY culture, then added 5% methanol every day to induce its expression. Then we verified the production of adhesive proteins by SDS-PAGE electrophoresis. However, we did not observe any bands in gel electrophoresis results, which means no protein is synthesized.
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[[File:T--BEIJING 4ELEVEN--AOX151_2.png|600px|thumb|center|Figure 2. SDS-PAGE gel analysis of supernatant samples during fp1-mfp5-fp1 fermentation]]
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Revision as of 07:13, 25 October 2020


AOX1 Promoter_fp1_mfp5_fp1_AOX1 Terminator

The circuit we transformed into Pichia Pastoris to produce fp151 protein.

Usage and Biology

Figure 1. Part demonstration

Our AOX1-fp1-mfp5-fp1 is a composite part consisting of an AOX1 Promoter, an fp1-mfp5-fp1 recombinant protein sequence, and an AOX1 Terminator. It regulates the expression of our recombinant adhesive protein. We inserted the sequence of the system onto vector pPIC9K and transferred the resulting plasmid into Pichia pastoris inoculated on BMMY culture, then added 5% methanol every day to induce its expression. Then we verified the production of adhesive proteins by SDS-PAGE electrophoresis. However, we did not observe any bands in gel electrophoresis results, which means no protein is synthesized.

Figure 2. SDS-PAGE gel analysis of supernatant samples during fp1-mfp5-fp1 fermentation


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 1896
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 937
    Illegal XhoI site found at 1191
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1962
  • 1000
    COMPATIBLE WITH RFC[1000]