Difference between revisions of "Part:BBa K4275002:Design"

 
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===Design Notes===
 
===Design Notes===
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1. The catalytic domain and the dockerin domain is interspaced with a glycine-rich linker.
  
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2. The 79 amino acid long type-I dockerin domain is fused at the terminal of the GS linker, followed by a TEV site and a 8xhis affinity purification tag (HHHHHHHH). 
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3. DNA sequence is codon-optimized based on the codon-usage table of <i>E.coli</i> Strain K12.MG1655, but the protein was later expressed in <i>K.marxianus</i> due to the lack of post-translational modifications and the formation of inclusion bodies in <i>E.coli</i> BL21(DE3).
  
 
===Source===
 
===Source===
  
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<i>Neocallimastix patriciarum</i> W5
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<br>
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<i>Clostridium thermocellum</i> (Dockerin-I domain)
  
 
===References===
 
===References===
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1.Chen, Hsin-Liang et al. "A Highly Efficient Β-Glucosidase From The Buffalo Rumen Fungus Neocallimastix Patriciarum W5". Biotechnology For Biofuels, vol 5, no. 1, 2012. Springer Science And Business Media LLC, https://doi.org/10.1186/1754-6834-5-24.

Latest revision as of 01:06, 14 October 2022


NpaBGS-t


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 502
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 1577
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1074
    Illegal AgeI site found at 88
    Illegal AgeI site found at 106
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

1. The catalytic domain and the dockerin domain is interspaced with a glycine-rich linker.

2. The 79 amino acid long type-I dockerin domain is fused at the terminal of the GS linker, followed by a TEV site and a 8xhis affinity purification tag (HHHHHHHH).

3. DNA sequence is codon-optimized based on the codon-usage table of E.coli Strain K12.MG1655, but the protein was later expressed in K.marxianus due to the lack of post-translational modifications and the formation of inclusion bodies in E.coli BL21(DE3).

Source

Neocallimastix patriciarum W5
Clostridium thermocellum (Dockerin-I domain)

References

1.Chen, Hsin-Liang et al. "A Highly Efficient Β-Glucosidase From The Buffalo Rumen Fungus Neocallimastix Patriciarum W5". Biotechnology For Biofuels, vol 5, no. 1, 2012. Springer Science And Business Media LLC, https://doi.org/10.1186/1754-6834-5-24.