Difference between revisions of "Part:BBa K3239009"

Revision as of 06:33, 18 October 2019

pMIT1-PRM1

PRM1 gene expressed by the P_MIT1 promoter.

Usage and Biology

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.

Unlike in the wildtype 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.

Figure 1. Illustration of the pGMP1-P_AOX1-GFP construct (for in trans regulations among homogeneous transcription factors and the homogenous AOX1, see the P_AOX1-GFP construct in part BBa_K3239007)

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal EcoRI site found at 2034
Illegal SpeI site found at 793
Illegal PstI site found at 2556
12
INCOMPATIBLE 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
21
INCOMPATIBLE 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
23
INCOMPATIBLE WITH RFC[23]
Illegal EcoRI site found at 2034
Illegal SpeI site found at 793
Illegal PstI site found at 2556
25
INCOMPATIBLE WITH RFC[25]
Illegal EcoRI site found at 2034
Illegal SpeI site found at 793
Illegal PstI site found at 2556
1000
COMPATIBLE WITH RFC[1000]

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K3239009 short</partinfo>
-PRM1 gene expressed by the pMIT1 promoter.
+''PRM1'' gene expressed by the ''P<sub>MIT1'' promoter.
 ===Usage and Biology===
-Upon methanol induction, pPRM1 expression of the homogeneous Prm1 is activated, and further amplified via the self-upregulation of Prm1. Prm1 then activates pMIT1, upregulating the expression of the homogeneous Mit1 and the heterogeneous Prm1.
+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.
-Unlike wildtype P. pastoris GS115 where Prm1 expression is then inhibited by Mit1, in the constructed pGMP1-pAOX1-GFP strain, the heterogeneous Prm1 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 up-regulate pAOX1 activity compared to wildtype P. pastoris.
+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''.
-Given that in constructed strains, pAOX1 not only expresses the yEGFP3 reporter gene, but also the homogenous AOX1 (alcohol oxidase 1) gene, the pGMP1-pAOX1-GFP strain should not only have a higher GFP yield compared to the pAOX1-GFP strain (pAOX1-GFP homologously recombined into wildtype P. pastoris GS115) but also exhibit higher growth rates since it should be more capable of metabolizing methanol.
+Given that in constructed strains, ''P<sub>AOX1'' not only expresses the ''yEGFP3'' reporter gene, but also the homogeneous ''AOX1'' gene, the ''pGMP1''-''P<sub>AOX1''-GFP strain should not only have a higher GFP yield compared to the ''P<sub>AOX1''-GFP strain (see part BBa_K3239007) but also exhibit higher growth rates since it should be more capable of metabolizing methanol.
-[[File:pGMP1-pAOX1-GFP.png|400px|thumb|Figure 1. Illustration of the pGMP1-pAOX1-GFP construct (for in trans regulations among homogenous transcription factors and the homogenous AOX1, see the pAOX1-GFP construct)]]
+[[File:pGMP1-pAOX1-GFP.png|400px|thumb|Figure 1. Illustration of the pGMP1-''P<sub>AOX1''-GFP construct (for in trans regulations among homogeneous transcription factors and the homogenous AOX1, see the ''P<sub>AOX1''-GFP construct in part BBa_K3239007)]]