Difference between revisions of "Part:BBa K3239009:Experience"

Latest revision as of 06:59, 18 October 2019

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Applications of BBa_K3239009

This construct is designed to moderately up-regulate the AOX1 promoter (P_AOX1) activity in P. pastoris GS115. It is to be integrated into the P_AOX1-GFP strain.

Upon methanol induction, P_PRM1 expression of the homogeneous PRM1 gene is activated, and further amplified via the self-upregulation of Prm1. Prm1 then activates P_MIT1, upregulating the expression of the homogeneous MIT1 gene and the heterogeneous PRM1 gene. This overall leads to a strong activation of P_AOX1 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-P_AOX1-GFP strain, the heterogeneous PRM1 gene is constantly self-upregulated due to the upregulation of P_MIT1 by Prm1. This leads to an overall upregulation of the expression of P_AOX1 transcription factors and hence shall further up-regulate P_AOX1 activity compared to wildtype P. pastoris GS115.

Given that in constructed strains, P_AOX1 not only expresses the yEGFP3 reporter gene, but also the homogeneous AOX1 gene, the pGMP1-P_AOX1-GFP strain should not only have a higher GFP yield compared to the P_AOX1-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

UNIQ1dba318f29789069-partinfo-00000000-QINU UNIQ1dba318f29789069-partinfo-00000001-QINU

@@ Line 5: / Line 5: @@
 ===Applications of BBa_K3239009===
-This construct is designed to moderately up-regulate the ''AOX1'' promoter (''P<sub>AOX1'') activity in ''P. pastoris GS115''. It is to be homogeneously recombined into the ''P<sub>AOX1''-GFP strain.
+This construct is designed to moderately up-regulate the ''AOX1'' promoter (''P<sub>AOX1'') activity in ''P. pastoris GS115''. It is to be integrated into the ''P<sub>AOX1''-GFP strain.
-Upon methanol induction, ''P<sub>PRM1'' expression of the homogeneous ''PRM1'' gene is activated, and further amplified via the self-upregulation of Prm1. Prm1 then activates ''P<sub>MIT1'', upregulating the expression of the homogeneous ''MIT1 gene and the heterogeneous ''PRM1'' gene.
+Upon methanol induction, ''P<sub>PRM1'' expression of the homogeneous ''PRM1'' gene is activated, and further amplified via the self-upregulation of Prm1. Prm1 then activates ''P<sub>MIT1'', upregulating the expression of the homogeneous ''MIT1'' gene and the heterogeneous ''PRM1'' gene. This overall leads to a strong activation of ''P<sub>AOX1'' 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''-''P<sub>AOX1''-GFP strain, the heterogeneous ''PRM1'' gene is constantly self-upregulated due to the upregulation of ''P<sub>MIT1'' by Prm1. This leads to an overall upregulation of the expression of ''P<sub>AOX1'' transcription factors and hence shall further up-regulate ''P<sub>AOX1'' activity compared to wildtype ''P. pastoris GS115''.