Difference between revisions of "Part:BBa K3598009"
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===Experiments and Results===
 
===Experiments and Results===
  
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[[File:T--BEIJING 4ELEVEN--tri_1.png|600px|thumb|center|Figure 1. Part demonstration]]
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[[File:T--BEIJING 4ELEVEN--tri_1.png|400px|thumb|center|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.
 
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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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.  
 
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]]
 
[[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]]
 
[[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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The results (Figure 5 & 6）indicate that tridecapeptide's efficiency to E. coli MG1655 were not significant, only showed obverse peak at 8 h. Its ability to kill P. acnes was relatively obvious at concentration 16mg/L, the final OD600 was about 14.6% decreased.
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K3598009 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3598009 SequenceAndFeatures</partinfo>

Revision as of 08:39, 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.

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

The results (Figure 5 & 6）indicate that tridecapeptide's efficiency to E. coli MG1655 were not significant, only showed obverse peak at 8 h. Its ability to kill P. acnes was relatively obvious at concentration 16mg/L, the final OD600 was about 14.6% decreased.