Coding

Part:BBa_K2865001:Design

Designed by: Chuqi Wang   Group: iGEM18_SMMU-China   (2018-09-29)
Revision as of 12:52, 12 October 2018 by WendyCui (Talk | contribs)


AR185-T2A-EGFP, nanobody inhibiting RyR2 phosphorylation


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 155
    Illegal NgoMIV site found at 302
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

After utilizing RyR2 to stimulate camels’ immune system and isolating the mRNA coding for heavy-chain antibodies, we got a gene library of camelids-S2808 nanobodies containing several million clones by reverse transcription and polymerase chain reaction. As for the screening techniques we chose phage display, a technique inserting the gene encoding S2808’s nanobody into a phage coat protein gene, causing the phage to "display" the nanobody on its outside while containing the gene for the protein on its inside. We transfected the phages into engineering bacteria and gained amplification products. Followed with multiple rounds of antigen affinity adsorption-elution-amplification, we finally got the S2808’s specific nanobody clones.