Part:BBa_K2262019

T7 Promoter+RBS+1095.6

Figure 1. P_T7+RBS+M-C+terminator

Introduction

      1095.6 is an peptide that can induces vasodilation in mice and rabbits. [1]. The score of SCM analysis shows its promising potential to be antifungal.

Scoring Card Predict

      1095.6 got 365.8 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 1095.6 has a high probability to have an antifungal function.

Figure 2. The sequence of Pe-4.
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 1095.6 has no different with negative control. It means the peptide has no obvious function on affecting the growth of mycelium.

Figure 3. The result shows that 1095.6 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 1095.6 with a concentration of 1000 μg/mL is higher than adding the negative control, which means the spore germination is inhibited by peptides and proves the peptides is effective. We can get the same result from adding 1095.6 with a concentration of 500 μg/mL and 250 μg/mL .

Figure 4. The result shows that 105.6 will affect the germination of spore.

3. Botany Experiment

      In order to test our anti-fungal peptides, we put 1095.6 and negative control, double-distilled water, on the flower. The negative control is on the left and 1095.6 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 1095.6, is not infected. It means 1095.6 has an anti-fungal function in real plant organism.

Figure 5. The result shows that 1095.6 will affect fungal infection.

Reference

[1] Vanessa Rioli; Benedito C. Prezoto; Katsuhiro Konno; Robson L. Melo; Clécio F. Klitzke; Emer S. Ferro; Mônica Ferreira-Lopes; Antonio C. M. Camargo; Fernanda C. V. Portaro. “A novel bradykinin potentiating peptide isolated from Bothrops jararacussu venom using catallytically inactive oligopeptidase EP24.15” 2008 Apr

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]