Part:BBa_K1022111
PT7: RBS: Signiferin: His 6
This Biobrick codes for promoter pT7 followed by Signiferin peptide with a His 6 tag.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Characterization
For more info, visit [http://2013.igem.org/Team:TU-Delft/PeptideCharacterization TU Delft iGEM13 Wiki]
Peptide Characterization:
An important part of the project is inhibition of growth or killing of bacteria with the use of antimicrobial peptides (AMPs). In order to get an idea of the toxicity of our peptides we conducted several minimal inhibiting concentration (MIC) experiments. These MIC measurements where done on E. coli, B. subtilis and S. delphini, with the first as representative of our Gram-negative expression host, the second for the Gram-positive targets and the last for our specific target.
MIC determination:
The MIC of signiferin on S. delphini, B. subtilis and E. coli were done according to the protocol described [http://2013.igem.org/Team:TU-Delft/PeptideCharacterization here]. S. delphini is the most sensitive for signiferin, as the MIC was determined to be 1µM (Figure 1A), no growth is seen at this concentration. The MIC for B. subtilis was determined to be 10µM (Figure 1B).
The expected selectivity of the chosen peptides for Staphylococcus was confirmed by these experiments, as for all the peptides for which we could determine a MIC, that MIC was the lowest on S. delphini.
For MIC by other peptides used in iGEM 2013 Team TU Delft, Click here.
Comparison:
The fact that for none of the peptides a MIC could be determined for E. coli (>150µM) further confirms the expected selectivity towards Gram-positives which is observed in the following graph(Figure 2):
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