Part:BBa_K3598012
AOX1 Promoter_CsgA_mfp5_mfp5_AOX1 Terminator
The circuit we transformed into Pichia Pastoris to produce AMP CEN1HC-Br.
Demonstration
Experiments and results
This part is a composite part consisitiong an AOX1 promter, CsgA-mfp5-mfp5 recombinant protein sequence, and an AOX1 terminator. The coding sequence of CsgA-mfp5-mfp5 has been codon optimized
Our AOX1-csgA-mfp5-mfp5 is a composite part consisting of an AOX1 Promoter, an csgA-mfp5-mfp5 recombinant protein sequence, and an AOX1 Terminator. It regulates the expression of our recombinant cohesive 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, the bands in the results are larger than they should be. This might be because that plasmids are integrated into the yeast genome with multiple copies, producing proteins that fold together.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 2127
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 937
Illegal XhoI site found at 1191 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 2193
- 1000COMPATIBLE WITH RFC[1000]
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