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 byhuman cytotoxic T lymphocyte and natural killer cells.

Figure 1. Part demonstration.

Usage and Biology

The GDP20 coding part was the original sequence of GDP20 we sent to a synthetic company to produce our GDP20 AMP. We verified the antimicrobial potency of GDP20 purchased from the company by adding its solution and placing pieces of soaked paper with its solution to plates inoculated with P. acnes and E. coli MG1655. The results showed inhibition zones in areas of direct contact between bacteria and GDP20, which means GDP20 is effective in killing both bacteria, and 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 GDP 20's ability to kill 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 indicate that GDP20's ability to kill E. coli MG1655 peaked at concentration 16mg/L, 8h, its effect reaching an OD600 of 2.0. GDP20's ability to kill P. acnes peaked at concentration 16mg/L, its effect reaching an OD600 of about 0.175.

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]