Difference between revisions of "Part:BBa K1462210:Design"
 
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===Design Notes===
 
===Design Notes===
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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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The n-butanol pathway in the cytoplasm is constructed as a control group to identify the effectiveness of the pathway directed to the mitochondria.Hbd,originated from Clostridium beijerinckii, expresses 3-hydroxybutyryl-CoA dehydrogenase, which converts AcAcCoA to HbCoA, the second step of the n-butanol pathway.This part is constructed in pUC57 individually at the beginning and it is assembled with Erg10(BBa_K1462210),Crt(BBa_K1462230) in plasmid 181 ,which is leucine auxotroph.
 
[[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.]]
 
[[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.]]
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===Source===
 
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S. cerevisiae genomic sequence
 
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===References===
 
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Eric J Steen, et al. (2008). Metabolic engineering of Saccharomyces cerevisiae for theproduction of n-butanol, Microbial Cell Factories, 7:36.
 

Latest revision as of 19:31, 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

The n-butanol pathway in the cytoplasm is constructed as a control group to identify the effectiveness of the pathway directed to the mitochondria.Hbd,originated from Clostridium beijerinckii, expresses 3-hydroxybutyryl-CoA dehydrogenase, which converts AcAcCoA to HbCoA, the second step of the n-butanol pathway.This part is constructed in pUC57 individually at the beginning and it is assembled with Erg10(BBa_K1462210),Crt(BBa_K1462230) in plasmid 181 ,which is leucine auxotroph.

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