Composite

Part:BBa_K2262017

Designed by: CHEN, PENG-TZU   Group: iGEM17_NCTU_Formosa   (2017-10-31)


T7 Promoter+RBS+M-C




Figure 1. PT7+RBS+M-C+terminator



Introduction


      M-C is a mast cell degranulating peptide [1]. The score of SCM analysis shows its promising potential to be antifungal.


Scoring Card Predict

      M-C got 387.08 from the Antifungal Scoring Card of Parabase System. The score is over 353, the threshold to divide whether a peptide has an antifungal function probability, that means M-C has a high probability to have an antifungal function.

Figure 2. The sequence of M-C.
The colors of each amino acids represent the probability of anti-fungal dipeptides.



Experiment


1. Inhibition Zone

      The result shows that the place we put M-C does not have little different with negative control. We cannot definitely sure if M-C can inhibit the growth of mycelium.


Figure 3. The result shows that M-C has no obvious function on affecting the growth of mycelium.


2. Spore Germination

      The result shows that the spore germination percentage of non-germinating spores adding M-C with a concentration of 1000 μg/mL was higher than adding the negative control, which means the spore germination is inhibited by peptides and proves the peptides are effective. We can get the same result from adding M-C with a concentration of 500 μg/mL and 250 μg/mL .


Figure 4. The result shows that M-C will affect the germination of spore.



3. Botany Experiment

      In order to test our anti-fungal peptides, we put M-C and negative control, double-distilled water, on the flower. The negative control is on the left and M-C is on the right. Both sides are infected with Botrytis cinerea. We checked the flower 3 days later whether the flower was affected or not. The result shows that the right side, which was spread on M-C, is not infected. It means M-C has an anti-fungal function in real plant organism.


Figure 5. The result shows that M-C will affect fungal infection.




Reference

[1] Argiolas A; Pisano JJ. “Isolation and characterization of two new peptides, mastoparan C and crabrolin, from the venom of the European hornet, Vespa crabro.” J Biol Chem. 1984 Aug 25;259(16):10106-11.

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
    COMPATIBLE WITH RFC[1000]


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