Difference between revisions of "Part:BBa K3598007"
 
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An antimicrobial peptide named CEN1HC-Br, it is isolated and characterized from the green sea urchin and shows broad-spectrum antibacterial activity.
 
An antimicrobial peptide named CEN1HC-Br, it is isolated and characterized from the green sea urchin and shows broad-spectrum antibacterial activity.
  
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===Usage and Biology===
 
  
 
[[File:T--BEIJING 4ELEVEN--CEN_1.png|400px|thumb|center|Figure 1. Part demonstration]]
 
[[File:T--BEIJING 4ELEVEN--CEN_1.png|400px|thumb|center|Figure 1. Part demonstration]]
  
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The CEN1HC-Br coding part was the original sequence of CEN1HC-Br we sent to a synthetic company to produce our CEN1HC-Br AMP. We verified the antimicrobial potency of CEN1HC-Br 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 CEN1HC-Br, which means CEN1HC-Br is effective in killing both bacteria, and would not affect bacteria outside its contact.  
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===Experiments and Results===
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The part is the original sequence of CEN1HC-Br. 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--Collection_Figure_2.png|400px|thumb|center|Figure 2. AMPs test methods]]
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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 the AMP, that indicates CEN1HC-Br possess the potency in killing both E.coli and P.acnes, while would not affect bacteria outside its contact.
 +
 
 
[[File:T--BEIJING 4ELEVEN--CEN3.png|400px|thumb|center|Figure 2. Plate verification result of CEN1HC-Br's ability to kill E. coli]]
 
[[File:T--BEIJING 4ELEVEN--CEN3.png|400px|thumb|center|Figure 2. Plate verification result of CEN1HC-Br's ability to kill E. coli]]
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[[File:T--BEIJING 4ELEVEN--CEN4.png|400px|thumb|center|Figure 3. Plate verification result of CEN1HC-Br's ability to kill P. acnes]]
 
[[File:T--BEIJING 4ELEVEN--CEN4.png|400px|thumb|center|Figure 3. Plate verification result of CEN1HC-Br's ability to kill P. acnes]]
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Then, we verified CEN1HC-Br'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 as a group, 0, 20, 24, 28, 32mg/L as a group, 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 CEN1HC-Br's ability to kill E. coli MG1655 peaked at concentrations 24 and 28 mg/L, at 6h and 24h, its effects reaching OD600 of about 0.85 and 1.25. CEN1HC-Br's ability to kill P. acnes peaked at concentration 16mg/L, its effect reaching an OD600 of about 1.8.  
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====OD600 Verification====
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Then, we verified CEN1HC-Br's ability against E. coli MG1655 and P. acnes by adding its solution to liquid medium inoculated with the two bacteria.  
 +
 
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We first test the efficiency to E.coli, the AMP concentration gradient as 0, 2, 4, 8, 12, 16 mg/L as a group, 0, 20, 24, 28, 32mg/L as a group, and measured the OD600 of cells through time. In the results (Figure.5), we can observe that the cell growth was almost not affected by CEN1HC-Br, especially at low concentration. When the concentration increased to 32 mg/mL, the growth rate of E.coli is only affected a little, but the final OD600 value of all groups at 24h is almost the same.
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[[File:T--BEIJING 4ELEVEN--CEN_4.png|400px|thumb|center|Figure 4. OD600 verification result of CEN1HC-Br's ability to kill E. coli]]
 
[[File:T--BEIJING 4ELEVEN--CEN_4.png|400px|thumb|center|Figure 4. OD600 verification result of CEN1HC-Br's ability to kill E. coli]]
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Since P. acnes has a slow growth rate, we only measured OD600 after 48 hours of cultivation. The results indicate that CEN1HC-Br's ability to kill P. acnes peaked at concentration 16 mg/L, its effect reaching OD600 of 0.19, about 26.9% less than the control 0.26.
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[[File:T--BEIJING 4ELEVEN--CEN_5.png|400px|thumb|center|Figure 5. OD600 verification result of CEN1HC-Br's ability to kill P. acnes]]
 
[[File:T--BEIJING 4ELEVEN--CEN_5.png|400px|thumb|center|Figure 5. OD600 verification result of CEN1HC-Br's ability to kill P. acnes]]
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Latest revision as of 17:23, 26 October 2020


CEN1HC-Br


An antimicrobial peptide named CEN1HC-Br, it is isolated and characterized from the green sea urchin and shows broad-spectrum antibacterial activity.


Figure 1. Part demonstration

Experiments and Results

The part is the original sequence of CEN1HC-Br. 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 the AMP, that indicates CEN1HC-Br possess the potency in killing both E.coli and P.acnes, while would not affect bacteria outside its contact.

Figure 2. Plate verification result of CEN1HC-Br's ability to kill E. coli
Figure 3. Plate verification result of CEN1HC-Br's ability to kill P. acnes

OD600 Verification

Then, we verified CEN1HC-Br's ability against E. coli MG1655 and P. acnes by adding its solution to liquid medium inoculated with the two bacteria.

We first test the efficiency to E.coli, the AMP concentration gradient as 0, 2, 4, 8, 12, 16 mg/L as a group, 0, 20, 24, 28, 32mg/L as a group, and measured the OD600 of cells through time. In the results (Figure.5), we can observe that the cell growth was almost not affected by CEN1HC-Br, especially at low concentration. When the concentration increased to 32 mg/mL, the growth rate of E.coli is only affected a little, but the final OD600 value of all groups at 24h is almost the same.

Figure 4. OD600 verification result of CEN1HC-Br's ability to kill E. coli

Since P. acnes has a slow growth rate, we only measured OD600 after 48 hours of cultivation. The results indicate that CEN1HC-Br's ability to kill P. acnes peaked at concentration 16 mg/L, its effect reaching OD600 of 0.19, about 26.9% less than the control 0.26.

Figure 5. OD600 verification result of CEN1HC-Br'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]