Coding

Part:BBa_K2114998:Design

Designed by: Wladislaw Stroukov   Group: iGEM16_Freiburg   (2016-10-05)
Revision as of 00:52, 7 October 2016 by WladStr (Talk | contribs) (→‎References)


pelB_aGFPnano


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 87
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 54
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

To ensure the formation of the characteristic disulfide bond of the nanobody the pelB leader sequence was included in the N-terminus of the nanobody by primer extensions and Gibson assembly.


Source

The anti-GFP nanobody is derived camelid heavy-chain antibodies and was provided by Dr. Maximilian Ulbrich (BIOSS).

References

1. Hamers-Casterman, C. et al. Naturally occurring antibodies devoid of light chains. Nature 363, 446–8 (1993).
2. Muyldermans, S. & Lauwereys, M. Unique single-domain antigen binding fragments derived from naturally occurring camel heavy-chain antibodies. J. Mol. Recognit. 12, 131–140 (1999).


Kubala MH, Kovtun O, Alexandrov K, Collins BM. (2010) Structural and thermodynamic analysis of the GFP:GFP-nanobody complex. Protein Sci; 19, 2389-401