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. 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]