Difference between revisions of "Part:BBa K1462210:Design"
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===Design Notes===
 
===Design Notes===
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The n-butanol pathway in the cytoplasm is constructed as a control group to identify the effectiveness of the pathway directed to the mitochondria. Erg10,native in Saccharomyces cerevisiae, expresses thiolase, which converts AcCoA to AcAcCoA, the first step of the n-butanol pathway.This part is constructed in pUC57 individually at the beginning and it is assembled with Hbd(BBa_K1462220),Crt(BBa_K1462230) in plasmid 181 ,which is leucine auxotroph.
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This part is constructed as a control group to identify our enzymes have been directed to the mitochondria.The GFP is fused at the C-terminus of Erg10 to report where the enzyme located.Under the confocal microscope, we can observe the location of green fluorescence, thus to confirm the exact subcellular localization of target proteins.
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[[File:Pathway.png|200px|left|frame|Figure 1. The n-butanol biosynthetic pathway. Enzymes are from these organisms: Erg10, S. cerevisiae; Hbd, Crt, AdhE2, Clostridium beijerinckii; Ccr, Streptomyces collinus.]]
  
 
===Source===
 
===Source===

Revision as of 09:04, 17 October 2014

Gal1+Erg10+cyc1


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1252
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 150
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This part is constructed as a control group to identify our enzymes have been directed to the mitochondria.The GFP is fused at the C-terminus of Erg10 to report where the enzyme located.Under the confocal microscope, we can observe the location of green fluorescence, thus to confirm the exact subcellular localization of target proteins.

Figure 1. The n-butanol biosynthetic pathway. Enzymes are from these organisms: Erg10, S. cerevisiae; Hbd, Crt, AdhE2, Clostridium beijerinckii; Ccr, Streptomyces collinus.

Source

S. cerevisiae genomic sequence

References

Eric J Steen, et al. (2008). Metabolic engineering of Saccharomyces cerevisiae for theproduction of n-butanol, Microbial Cell Factories, 7:36.