Difference between revisions of "Part:BBa K3239009"
Nicole Guo (Talk | contribs) (→Usage and Biology) |
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===Usage and Biology=== | ===Usage and Biology=== | ||
− | 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'' of the ''AOX1'' gene. |
Unlike in the wildtype ''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''. | Unlike in the wildtype ''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''. |
Revision as of 06:45, 18 October 2019
pMIT1-PRM1
PRM1 gene expressed by the PMIT1 promoter.
Usage and Biology
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 of the AOX1 gene.
Unlike in the wildtype 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.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 2034
Illegal SpeI site found at 793
Illegal PstI site found at 2556 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 2034
Illegal NheI site found at 3815
Illegal SpeI site found at 793
Illegal PstI site found at 2556 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 2034
Illegal BglII site found at 1387
Illegal BamHI site found at 2917
Illegal BamHI site found at 3836 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 2034
Illegal SpeI site found at 793
Illegal PstI site found at 2556 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 2034
Illegal SpeI site found at 793
Illegal PstI site found at 2556 - 1000COMPATIBLE WITH RFC[1000]