Part:BBa_K3239009:Experience
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Applications of BBa_K3239009
This construct is designed to moderately up-regulate the AOX1 promoter (PAOX1) activity in P. pastoris GS115. It is to be integrated into the PAOX1-GFP strain.
Upon methanol induction, PPRM1 expression of the homogeneous PRM1 gene is activated, and further amplified via the self-upregulation of Prm1. Prm1 then activates PMIT1, upregulating the expression of the homogeneous MIT1 gene and the heterogeneous PRM1 gene. This overall leads to a strong activation of PAOX1 expression of the AOX1 gene.
Unlike in the wild type P. pastoris GS115 where PRM1 gene expression is then inhibited by Mit1, in the pGMP1-PAOX1-GFP strain, the heterogeneous PRM1 gene is constantly self-upregulated due to the upregulation of PMIT1 by Prm1. This leads to an overall upregulation of the expression of PAOX1 transcription factors and hence shall further up-regulate PAOX1 activity compared to wildtype P. pastoris GS115.
Given that in constructed strains, PAOX1 not only expresses the yEGFP3 reporter gene, but also the homogeneous AOX1 gene, the pGMP1-PAOX1-GFP strain should not only have a higher GFP yield compared to the PAOX1-GFP strain (see part BBa_K3239007) but also exhibit higher growth rates since it should be more capable of metabolizing methanol.
User Reviews
For detailed modeling, design, experiment, results, and demonstration, please check our wiki:
https://2019.igem.org/Team:ShanghaiFLS_China/Model
https://2019.igem.org/Team:ShanghaiFLS_China/Design
https://2019.igem.org/Team:ShanghaiFLS_China/Experiments
https://2019.igem.org/Team:ShanghaiFLS_China/Results
https://2019.igem.org/Team:ShanghaiFLS_China/Demonstrate
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