Difference between revisions of "Part:BBa K3598009"
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A short antimicrobial peptide containing 13 amino acids.
 
A short antimicrobial peptide containing 13 amino acids.
  
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===Usage and Biology===
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===Experiments and Results===
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[[File:T--BEIJING 4ELEVEN--tri_1.png|600px|thumb|center|Figure 1. Part demonstration]]
 
[[File:T--BEIJING 4ELEVEN--tri_1.png|600px|thumb|center|Figure 1. Part demonstration]]
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The tridecapeptide coding part was the original sequence of tridecapeptide we sent to a synthetic company to produce our tridecapeptide AMP. We verified the antimicrobial potency of tridecapeptide purchased from the company by adding its solution and placing pieces of paper soaked 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 tridecapeptide, which means tridecapeptide is effective in killing both bacteria, and would not affect bacteria outside its contact.  
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[[File:T--BEIJING 4ELEVEN--tri_2.png|600px|thumb|center|Figure 2. Plate verification result of tridecapeptide's ability to kill E. coli]]
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The tridecapeptide coding part is the original sequence of tridecapeptide, which is a quite short 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.
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[[File:T--BEIJING 4ELEVEN--tri_3.png|600px|thumb|center|Figure 3. Plate verification result of tridecapeptide's ability to kill P. acnes]]
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Then, we verified tridecapeptide'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, 16mg/L, and measured the OD600 of the E. coli through time. Since P. acnes has a slow growth rate, we only measured OD600 after 24 hours of cultivation. The results indicate that tridecapeptide's ability to kill E. coli MG1655 peaked at concentration 16 mg/L, 8h, its effects reaching OD600 of about 1.8. CEN1HC-Br's ability to kill P. acnes peaked at concentration 16mg/L, its effect reaching an OD600 of about 0.24.  
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[[File:T--BEIJING_4ELEVEN--Collection_Figure_2.png|400px|thumb|center|Figure 2. AMPs test methods]]
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[[File:T--BEIJING 4ELEVEN--tri_4.png|600px|thumb|center|Figure 4. OD600 verification result of tridecapeptide's ability to kill E. coli]]
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[[File:T--BEIJING 4ELEVEN--tri_5.png|600px|thumb|center|Figure 5. OD600 verification result of tridecapeptide's ability to kill P. acnes]]
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====Plate verification Results====
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The plate results showed inhibition zones in areas of direct contact between bacteria and tridecapeptide, which means tridecapeptide is effective in killing both E.coli and P.acnes, while would not affect bacteria outside its contact. However, the edge of the zone of inhibition is not clear when test to E.coli MG655，which means tridecapeptide’s efficiency to E. coli is not significant. Additionally, the zone of inhibition in P.acnes plate was blurry, that may due to the hydrophobicity of this short peptide.
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[[File:T--BEIJING 4ELEVEN--tri_2.png|400px|thumb|center|Figure 3. Plate verification result of tridecapeptide's efficiency to E. coli MG1655]]
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[[File:T--BEIJING 4ELEVEN--tri_3.png|400px|thumb|center|Figure 4. Plate verification result of tridecapeptide's efficiency to P. acnes]]
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====OD600 Verification====
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Then, we verified tridecapeptide'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, 16mg/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.  
 +
 
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The results indicate that tridecapeptide's ability to kill E. coli MG1655 peaked at concentration 16 mg/L, 8h, its effects reaching OD600 of about 1.8. CEN1HC-Br's ability to kill P. acnes peaked at concentration 16mg/L, its effect reaching an OD600 of about 0.24.  
 +
 
 +
[[File:T--BEIJING 4ELEVEN--tri_4.png|400px|thumb|center|Figure 5. OD600 verification result of tridecapeptide's ability to kill E. coli]]
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[[File:T--BEIJING 4ELEVEN--tri_5.png|400px|thumb|center|Figure 6. OD600 verification result of tridecapeptide's ability to kill P. acnes]]
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Revision as of 08:28, 26 October 2020


tridecapeptide

A short antimicrobial peptide containing 13 amino acids.

Experiments and Results

Figure 1. Part demonstration

The tridecapeptide coding part is the original sequence of tridecapeptide, which is a quite short 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 showed inhibition zones in areas of direct contact between bacteria and tridecapeptide, which means tridecapeptide is effective in killing both E.coli and P.acnes, while would not affect bacteria outside its contact. However, the edge of the zone of inhibition is not clear when test to E.coli MG655，which means tridecapeptide’s efficiency to E. coli is not significant. Additionally, the zone of inhibition in P.acnes plate was blurry, that may due to the hydrophobicity of this short peptide.

Figure 3. Plate verification result of tridecapeptide's efficiency to E. coli MG1655
Figure 4. Plate verification result of tridecapeptide's efficiency to P. acnes

OD600 Verification

Then, we verified tridecapeptide'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, 16mg/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 tridecapeptide's ability to kill E. coli MG1655 peaked at concentration 16 mg/L, 8h, its effects reaching OD600 of about 1.8. CEN1HC-Br's ability to kill P. acnes peaked at concentration 16mg/L, its effect reaching an OD600 of about 0.24.

Figure 5. OD600 verification result of tridecapeptide's ability to kill E. coli
Figure 6. OD600 verification result of tridecapeptide'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]