Difference between revisions of "Part:BBa K3712021"
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===USAGE AND BIOLOGY=== | ===USAGE AND BIOLOGY=== | ||
| + | <p>BLP-7 is an antimicrobial peptide consisting of 27 amino acids and has been shown to have anti-inflammatory effects and a broad spectrum of antibacterial activity. </p> | ||
| + | <p>Cell experiments shows that BLP-7 inhibits the expression of two proinflammatory cytokines interleukin (IL)-8 and granulocyt-macrophage colony-stimulating factor (GM-CSF) in normal human epidermal keratinocytes (NHEKs). Mice experiments have also shown that BLP-7 can significantly inhibit skin inflammation caused by P.acnes. As most other antimicrobial peptides (AMPs), BLP-7 can target the lipid bilayer of bacterial cell plasma membrane, punch holes through the membrane and kill bacteria. BLP-7 dose not lead to antibiotic resistance and could be used as a potential anti-acne agent. </p> | ||
| + | <p>Methods:Bacteriostatic experiment one: inhibition zone experiment</p> | ||
| + | <p>preparing solid medium: Heat and melt the BHI medium, then cool it to about 50℃. Add 1 mL of bacterial suspension to every 100 mL medium. Shake to ensure that the bacteria distribute evenly and pour into a large culture dish. Place Oxford-cups on the surface of the plate corresponding to the marked number, and dropwise add antimicrobial peptide sample with concentration gradient to Oxford-cups. Incubate together for a period of time and measure the diameter of inhibition zone.</p> | ||
| + | <p>Bacteriostatic experiment two: absorbance method</p> | ||
| + | Add BLP-7 at concentrations ranging from 1 to 1000μM to the culture medium and incubate anaerobically at 37°C for 24 hours using antibiotics as positive control and sterile water as negative control. The MIC value of BLP-7 is clarified by measuring the absorbance at 630 nm using a microplate reader. Assume the concentration with no significant difference from the positive control group MIC.</p> | ||
| + | <p>Future direction:</p> | ||
| + | <p>(1) Due to the lack of experience and method of Propionibacterium acnes cultivation, we fail to ascertain the optimal growth condition of Propionibacterium acnes after a long period of exploration although we successfully culture it. Subsequent exploration will be carried out to find the appropriate method to control the growth of Propionibacterium acnes.</p> | ||
| + | <p>(2) After determining the optimal growth conditions, we will further explore the principles of antibacterial experiments due to the uncertainty of incubation time and temperature of antimicrobial peptides.</p> | ||
===RESULTS=== | ===RESULTS=== | ||
Revision as of 03:57, 21 October 2021
Wild-type Bombinin-like peptide 7 (BLP-7)
USAGE AND BIOLOGY
BLP-7 is an antimicrobial peptide consisting of 27 amino acids and has been shown to have anti-inflammatory effects and a broad spectrum of antibacterial activity.
Cell experiments shows that BLP-7 inhibits the expression of two proinflammatory cytokines interleukin (IL)-8 and granulocyt-macrophage colony-stimulating factor (GM-CSF) in normal human epidermal keratinocytes (NHEKs). Mice experiments have also shown that BLP-7 can significantly inhibit skin inflammation caused by P.acnes. As most other antimicrobial peptides (AMPs), BLP-7 can target the lipid bilayer of bacterial cell plasma membrane, punch holes through the membrane and kill bacteria. BLP-7 dose not lead to antibiotic resistance and could be used as a potential anti-acne agent.
Methods:Bacteriostatic experiment one: inhibition zone experiment
preparing solid medium: Heat and melt the BHI medium, then cool it to about 50℃. Add 1 mL of bacterial suspension to every 100 mL medium. Shake to ensure that the bacteria distribute evenly and pour into a large culture dish. Place Oxford-cups on the surface of the plate corresponding to the marked number, and dropwise add antimicrobial peptide sample with concentration gradient to Oxford-cups. Incubate together for a period of time and measure the diameter of inhibition zone.
Bacteriostatic experiment two: absorbance method
Add BLP-7 at concentrations ranging from 1 to 1000μM to the culture medium and incubate anaerobically at 37°C for 24 hours using antibiotics as positive control and sterile water as negative control. The MIC value of BLP-7 is clarified by measuring the absorbance at 630 nm using a microplate reader. Assume the concentration with no significant difference from the positive control group MIC.</p>
Future direction:
(1) Due to the lack of experience and method of Propionibacterium acnes cultivation, we fail to ascertain the optimal growth condition of Propionibacterium acnes after a long period of exploration although we successfully culture it. Subsequent exploration will be carried out to find the appropriate method to control the growth of Propionibacterium acnes.
(2) After determining the optimal growth conditions, we will further explore the principles of antibacterial experiments due to the uncertainty of incubation time and temperature of antimicrobial peptides.
RESULTS
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]