Difference between revisions of "Part:BBa K3239009"

Latest revision as of 06:47, 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. This overall leads to a strong activation of P_AOX1 expression of the AOX1 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. This overall leads to a strong activation of ''P<sub>AOX1'' expression of the ''AOX1'' 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)]]