Part:BBa_K3239009

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]