Difference between revisions of "Part:BBa K1489004:Design"

 
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===Design Notes===
 
===Design Notes===
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When designing this part, had to choose between the pBAD and pLac promoters. Decided to use the pBAD promoter due to tighter regulation; we were unsure of whether the protein was toxic and did not want to have too much leaky expression.
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Lpp-OmpA-linker (BBa_K103006) was chosen to serve as the anchor in the outer membrane due to its previous uses in biobricks containing protein assemblies anchored in the outer membrane.
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Bovine galectin was used as a model galectin. As a mammalian galectin, it is a smaller protein than CvGal1 (BBa_K1489005), the galectin found in the Eastern Oyster.
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When designing this part, we had to choose between the pBAD and pLac promoters. Decided to use the pBAD promoter due to believed tighter regulation; we were unsure of whether the protein was toxic and did not want to have too much leaky expression.
  
  
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===References===
 
===References===
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Bourne Y, Bolgiano B, Liao DI, et al. Crosslinking of mammalian lectin (galectin-1) by complex biantennary saccharides. Nat Struct Biol. 1994;1(12):863-70.

Latest revision as of 00:04, 17 October 2014

pBAD-RBS-OmpA-Bovine Galectin-1-B0012 Terminator


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 125
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 65
    Illegal XhoI site found at 971
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Lpp-OmpA-linker (BBa_K103006) was chosen to serve as the anchor in the outer membrane due to its previous uses in biobricks containing protein assemblies anchored in the outer membrane.

Bovine galectin was used as a model galectin. As a mammalian galectin, it is a smaller protein than CvGal1 (BBa_K1489005), the galectin found in the Eastern Oyster.

When designing this part, we had to choose between the pBAD and pLac promoters. Decided to use the pBAD promoter due to believed tighter regulation; we were unsure of whether the protein was toxic and did not want to have too much leaky expression.


Source

OmpA is from E. coli genome Bovine galectin 1 is from Bos taurus genome.

References

Bourne Y, Bolgiano B, Liao DI, et al. Crosslinking of mammalian lectin (galectin-1) by complex biantennary saccharides. Nat Struct Biol. 1994;1(12):863-70.