Composite

Part:BBa_K2262021

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


T7 Promoter+RBS+T-SN2




Figure 1. PT7+RBS+T-SN2+terminator



Introduction


      T-SN2 is an antimicrobial peptide [1]. The score of SCM analysis shows its promising potential to be antifungal.


Scoring Card Predict

      T-SN2 got 438.5 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 T-SN2 has a high probability to have an antifungal function.

Figure 2. The sequence of T-SN2.
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 T-SN2 will have a hole. It means the peptide will affect the growth of mycelium.


Figure 3. The result shows that T-SN2 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 T-SN2 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 T-SN2 with a concentration of 500 μg/mL and 250 μg/mL .


Figure 4. The result shows that T-SN2 will affect the germination of spore.



3. Botany Experiment

      In order to test our anti-fungal peptides, we put T-SN2 and negative control, double-distilled water, on the flower. The negative control is on the left and T-SN2 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 T-SN2 is not infected. It means T-SN2 has an anti-fungal function in real plant organism.


Figure 5. The result shows that T-SN2 will affect fungal infection.




Reference

[1] Xiaolong Yang; Yuhong Hu; Shiqi Xu; Yonghong Hu; Hao Meng; Chao Guo; Yuliang Liu; Jingze Liu; Zhijun Yu; Hui Wang. “Identification of multiple antimicrobial peptides from the skin of fine-spined frog, Hylarana spinulosa (Ranidae)” Biochimie Volume 95, Issue 12, December 2013, Pages 2429-2436


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]


[edit]
Categories
Parameters
None