Difference between revisions of "Part:BBa K4275002:Design"
(→‎Design Notes)
 
(One intermediate revision by the same user not shown)
Line 12: Line 12:
 
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).   
 
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 <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 E.Coli BL21(DE3).
+
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===
  
−
<i>Neocallimastix patriciarum</i> W5
+
<i>Neocallimastix patriciarum</i> W5  
 +
<br>
 
<i>Clostridium thermocellum</i> (Dockerin-I domain)
 
<i>Clostridium thermocellum</i> (Dockerin-I domain)
  

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.