DNA

Part:BBa_K3593002

Designed by: Yiming Dong   Group: iGEM20_GreatBay_SCIE   (2020-10-05)
Revision as of 04:20, 27 October 2020 by RWSZ (Talk | contribs)

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ssDNA, aptamer for α-amanitin(H06)

Aptamer of α-amatoxin found in another literature using Sepharose immobilized method of SELEX. Its function of binding is characterized by ELONA and dot blot, by which we repeat the result successfully.

Background

Amatoxins are chemicals present inside the genus Amanita and caused about 90% of mushroom poisoning. Being able to detect it before eating or in the field could possibly make a great decrease in people and animals who died because of poisonous mushrooms. Also being able to detect it in hospital can greatly help doctors in mushroom areas get correct information and do effective diagnosis to save the patient.
Aptamers are oligonucleotides that form secondary structures, giving them the ability to bind targeted molecules, including ions or small molecules, and, in our case, amanitin.

Sequence and features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 44

Characterisation of this part

Experiment data

BBa_K3593002 is an aptamer of α-amanitin named H06. It comes from a previous literature.[1] Its binding affinity is tested via a method called ELONA(enzyme-linked oligonucleotide assay) which is from the same essay,with a similar principle to ELISA. We also conduct ELONA on it by ourselves to test it's binding.

T--GreatBay SCIE--Fig.1 ELONA on H06 aptamer.png

Fig.1 ELONA on H06 aptamer.

References

1.Han Q, Xia X, Jing L, et al. Selection and characterization of DNA aptamer specially targeting α-amanitin in wild mushrooms. SDRP J Food Sci Technol. 2018;3(6):497-508.



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