Difference between revisions of "Part:BBa K3182009"

 
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<partinfo>BBa_K3182009 short</partinfo>
 
<partinfo>BBa_K3182009 short</partinfo>
  
This part is a de novo antimicrobial peptide. The antimicrobial peptide was generated by the Parseq model and the antimicrobial activity was predicted by the Parseq model. This part was designed to be assembled with BBa_K3182010.
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Parseq Alpha is a de novo antimicrobial peptide. It was generated by the Parseq model in combination with an evolution script.  This part was first ordered chemically synthesized by Caslo to investigate its antimicrobial activity on its own. The peptides were dissolved in 6 M Urea due to their hydrophobic nature. The activity was tested on <i>E.coli</i> and <i>B.Subtilis</i>. The antimicrobial activity assay was carried out in a 96 well plate. Bacteria were combined with the antimicrobial peptides in different concentrations. They were then incubated at 37°C, with shaking, every 15 minutes. The plate was incubated for 16 hours and OD<sub>600</sub> was measured. The results can be seen in <b>Figure 1.</b> Parseq Alpha showed an inhibiting effect on the growth of <i>E.coli</i>. A concentration of 30 µg/mL completely inhibited the growth of <i>E.coli</i>. In comparison, <i>B.subtilis</i> could not be inhibited completely by Parseq Alpha. However, high concentrations of the peptide showed partial inhibition of the growth of <i>B.subtilis</i>, where the lag phase is extended.  
  
  
 
[[File:T--Linkoping_Sweden--AlphaEcoli.png|650px|thumb|center|<b>Figure 1</b> Shows the antimicrobial activity of Parseq Alpha  
 
[[File:T--Linkoping_Sweden--AlphaEcoli.png|650px|thumb|center|<b>Figure 1</b> Shows the antimicrobial activity of Parseq Alpha  
  on <i>E.Coli.</i>. The OD<sup>600</sup> value is displayed on the y-axis and the concentrations tested can be seen in on the x-axis. NG stands for no growth and MIC stands for minimum inhibitory concentration. The positive control was E.coli cultured without antimicrobial peptides. The samples were cultured in 200 µL in a 96 well plate and incubated at 37°C for 16 hours. All measurements are done in 6 replicates. Error bars represent SD. ]]
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  on A) <i>E.Coli.</i> and B) <i>B.subtilis</i>. The samples were cultured in 200 µL low salt LB, in a 96 well plate and incubated at 37°C. Shaking was conducted for 1 minute every 15 minutes and the OD<sub>600</sub> was measured every hour for 16 hours. The negative control represents the bacterial growth with no antimicrobial peptides present but the same buffer the peptides were dissolved in. All measurements are done in 5 replicates and error bars represent SD. The graphs were created in Graphpad.]]
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[[File:T--Linkoping Sweden--alphaBsubtilis.png|650px|thumb|center|<b>Figure 1.</b> Shows the antimicrobial activity of Parseq Alpha on B.Subtilis. The OD<sup>600</sup> value is displayed on the y-axis and the concentrations tested can be seen in on the x-axis. The positive control was B.Subtilis cultured without antimicrobial peptides. The samples were cultured in 200 µL in a 96 well plate and incubated at 37°C for 16 hours. All measurements are done in 6 replicates. Error bars represent SD.]]
 
  
 
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<!-- Add more about the biology of this part here

Latest revision as of 17:02, 21 October 2019


Parseq-α

Parseq Alpha is a de novo antimicrobial peptide. It was generated by the Parseq model in combination with an evolution script. This part was first ordered chemically synthesized by Caslo to investigate its antimicrobial activity on its own. The peptides were dissolved in 6 M Urea due to their hydrophobic nature. The activity was tested on E.coli and B.Subtilis. The antimicrobial activity assay was carried out in a 96 well plate. Bacteria were combined with the antimicrobial peptides in different concentrations. They were then incubated at 37°C, with shaking, every 15 minutes. The plate was incubated for 16 hours and OD600 was measured. The results can be seen in Figure 1. Parseq Alpha showed an inhibiting effect on the growth of E.coli. A concentration of 30 µg/mL completely inhibited the growth of E.coli. In comparison, B.subtilis could not be inhibited completely by Parseq Alpha. However, high concentrations of the peptide showed partial inhibition of the growth of B.subtilis, where the lag phase is extended.


Figure 1 Shows the antimicrobial activity of Parseq Alpha on A) E.Coli. and B) B.subtilis. The samples were cultured in 200 µL low salt LB, in a 96 well plate and incubated at 37°C. Shaking was conducted for 1 minute every 15 minutes and the OD600 was measured every hour for 16 hours. The negative control represents the bacterial growth with no antimicrobial peptides present but the same buffer the peptides were dissolved in. All measurements are done in 5 replicates and error bars represent SD. The graphs were created in Graphpad.


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]