Part:BBa_K5415004
To test the functionality of our biosensors, we first needed to “humanise” the yeast by introducing the HIF-1 system, as it was pivotal for mimicking hypoxic cellular reactions, akin to those experienced in human physiology, which, in turn, were crucial for our biosensor assays. By utilising the GAL1,10 promoter, we ensured robust expression of the HIF-1 components, as this promoter is known for its strong activation in the presence of galactose, thereby facilitating the desired gene expression under specific conditions. In our approach, we utilised two key iGEM BioBricks: HIF-1 beta and HIF-1 alpha, by integrating which we are able to create a functional hypoxia signalling cascade that closely resembles that found in human cells. ADH1 and ADH2 terminators were deployed to ensure precise regulation of gene expression and prevention of any unintended transcriptional read-through.
The successful implementation of this humanised yeast model opens up new avenues for biosensor development and testing. By leveraging the HIF-1 system, this composite part could not only contribute to a deeper understanding of cellular responses to oxygen deprivation but would also pave the way for innovative applications in medical diagnostics and/or environmental monitoring.
HIF1 system for yeast humanisation.
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 3244
Illegal BglII site found at 3452
Illegal BamHI site found at 3355 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 929
Illegal NgoMIV site found at 3383
Illegal AgeI site found at 2972
Illegal AgeI site found at 4459 - 1000COMPATIBLE WITH RFC[1000]
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