Difference between revisions of "Part:BBa K3598006"
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<partinfo>BBa_K3598006 short</partinfo>
 
<partinfo>BBa_K3598006 short</partinfo>
  
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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.
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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 by human cytotoxic T lymphocyte and natural killer cells.
  
 
[[File:T--BEIJING 4ELEVEN--GDP20_demonstration.png|400px|thumb|center|Figure 1. Part demonstration.]]
 
[[File:T--BEIJING 4ELEVEN--GDP20_demonstration.png|400px|thumb|center|Figure 1. Part demonstration.]]
  
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===Usage and Biology===
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===Experiments and Results===
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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.  
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This part is the original sequence of GDP20,which is an excellent 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--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 of GDP20 are shown in Figure 3&4, from which we can see clear inhibition zones against both E.coli MG1655 and P.acnes. And the inhibiton areas are limited to the position that directly contacts the AMP. So we can infer that GDP20 is effective in killing both E.coli and P.acnes, while would not affect bacteria outside its contact.
  
 
[[File:T--BEIJING 4ELEVEN--GDP20_fig3 to E.Coli.png|400px|thumb|center|Figure 3. Plate verification result of GDP20's ability to kill E. coli]]
 
[[File:T--BEIJING 4ELEVEN--GDP20_fig3 to E.Coli.png|400px|thumb|center|Figure 3. Plate verification result of GDP20's ability to kill E. coli]]
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[[File:T--BEIJING 4ELEVEN--GDP20_fig4.png|400px|thumb|center|Figure 3. Plate verification result of GDP20's ability to kill P. acnes]]
 
[[File:T--BEIJING 4ELEVEN--GDP20_fig4.png|400px|thumb|center|Figure 3. Plate verification result of GDP20's ability to kill P. acnes]]
  
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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.  
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Then, we verified GDP20's ability against to 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.  
  
 
[[File:T--BEIJING 4ELEVEN--GDP20_fig5.png|400px|thumb|center|Figure 4. OD600 verification result of GDP20's ability to kill E. coli]]
 
[[File:T--BEIJING 4ELEVEN--GDP20_fig5.png|400px|thumb|center|Figure 4. OD600 verification result of GDP20's ability to kill E. coli]]

Revision as of 14:06, 26 October 2020


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 by human cytotoxic T lymphocyte and natural killer cells.

Figure 1. Part demonstration.

Experiments and Results

This part is the original sequence of GDP20,which is an excellent 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 of GDP20 are shown in Figure 3&4, from which we can see clear inhibition zones against both E.coli MG1655 and P.acnes. And the inhibiton areas are limited to the position that directly contacts the AMP. So we can infer that GDP20 is effective in killing both E.coli and P.acnes, while 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 GDP20's ability against to 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]