Part:BBa_K3598006

GDP20

Granulysin-derived peptides. Granulysin is a human AMP of the saposin-like family, a family of peptides that has been conserved from amoe-bas to humans, and a novel effector molecule released by human cytotoxic T lymphocyte and natural killer cells.

Figure 1. Part demonstration.

Experiments and Results

This part is the original sequence of GDP20,which is an excellent antimicrobial pepetide. We verified its antimicrobial potency through two methods: plate inhibition zones assay and OD600 analysis. In plate inhibition zones assay,wo placed pieces of paper soaked with AMP solutions onto plates inoculated with E.coli MG1655 and P.acnes and observing bacteria growth inhibition zones. In OD600 analysis, we added AMP solutions into liquid culture inoculated with the bacteria and measuring OD600 value.

Figure 2. AMPs test methods

Plate verification Results

The plate results of GDP20 are shown in Figure 3&4, from which we can see clear inhibition zones against both E.coli MG1655 and P.acnes. And the inhibiton areas are limited to the position that directly contacts the AMP. So we can infer that GDP20 is effective in killing both E.coli and P.acnes, while would not affect bacteria outside its contact.

Figure 3. Plate verification result of GDP20's ability to kill E. coli

Figure 3. Plate verification result of GDP20's ability to kill P. acnes

Then, we verified GDP20's ability against to E. coli MG1655 and P. acnes by adding its solution to liquid culture inoculated with the two bacteria. We set the AMP concentration gradient as 0, 2, 4, 8, 12，16 mg/L, and measured the OD600 of the E. coli through time. Since P. acnes has a slow growth rate, we only measured OD600 after 48 hours of cultivation. The results in Figure 5&6 indicate that GDP20's exhibit perfect potency agianst to the two bacteria, espesially to P.acnes. On the demostration to E.coli, GDP20 only has a significant effect at concentration 16mg/L. When against to P. acnes, the OD600 value of control group was 0.26 after 48 hours from inoculation, while the experimental group was 0.17, about 34.4% dropped.

Figure 4. OD600 verification result of GDP20's ability to kill E. coli

Figure 5. OD600 verification result of GDP20's ability to kill P. acnes

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]