Difference between revisions of "Part:BBa K4129002"
 
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<partinfo>BBa_K4129002 short</partinfo>
 
<partinfo>BBa_K4129002 short</partinfo>
  
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The part consists of the 1000 bp upstream of Formate Dehydrogenase (FDH) CDS from Aspergillus niger. Since FDH genes have been shown to be upregulated in response to furfural in Saccharomyces cerevisiae, the part was utilised to test if anFDH promoter is also responsive to furfural. When used to drive expression of mCherry in A. niger through transient expression, it is neither constitutive nor induced by furfural or does not promote expression at levels high enough to be detected during plasmid based expression.
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The part consists of the 1000 bp upstream of Formate Dehydrogenase (FDH) CDS from Aspergillus niger. Since FDH genes have been shown to be upregulated in response to furfural in Saccharomyces cerevisiae, the part was utilised to test if anFDH promoter is also responsive to furfural. Quantification by qPCR of the FDH transcript and mCherry transcript from a transiently expressed BBa_K4129005 (BBa_K4129002 regulating mCherry reporter), showed that BBa_K4129002 is induced by furfural both in its native regulatory enviroment (Fig. 1 A) and when used in syntetic expression cassettes (Fig 1 B). The promoter shows a considerable dynamic range in response to furfural, making it ideal for enginnering biosensors. To our knowledge, such a biosensor would be the first furfural biosensor ever built.
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<figure><img style="width: 80%; padding:28px;"src="https://static.igem.org/mediawiki/parts/0/0d/Registry_fig1.png" class="safetyfirstimg"><figcaption>Figure 1:  Investigation of anFDH promoter activity in response to furfural. The plots show the mRNA levels relative to actin mRNA of FDH (A) and mCherry transiently expressed from BBa_K4129005 (B) as measured by qPCR.</figcaption></figure>
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Latest revision as of 22:20, 13 October 2022


Promoter of the Aspergillus niger FDH gene

The part consists of the 1000 bp upstream of Formate Dehydrogenase (FDH) CDS from Aspergillus niger. Since FDH genes have been shown to be upregulated in response to furfural in Saccharomyces cerevisiae, the part was utilised to test if anFDH promoter is also responsive to furfural. Quantification by qPCR of the FDH transcript and mCherry transcript from a transiently expressed BBa_K4129005 (BBa_K4129002 regulating mCherry reporter), showed that BBa_K4129002 is induced by furfural both in its native regulatory enviroment (Fig. 1 A) and when used in syntetic expression cassettes (Fig 1 B). The promoter shows a considerable dynamic range in response to furfural, making it ideal for enginnering biosensors. To our knowledge, such a biosensor would be the first furfural biosensor ever built.

Figure 1: Investigation of anFDH promoter activity in response to furfural. The plots show the mRNA levels relative to actin mRNA of FDH (A) and mCherry transiently expressed from BBa_K4129005 (B) as measured by qPCR.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 34
    Illegal SpeI site found at 160
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal SpeI site found at 34
    Illegal SpeI site found at 160
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 34
    Illegal SpeI site found at 160
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 34
    Illegal SpeI site found at 160
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 564
    Illegal BsaI site found at 929