Difference between revisions of "Part:BBa K1022112"

Revision as of 13:00, 3 October 2013

PT7: RBS: Magainin II: His 6

This biobrick codes for the promoter pT7, Magainin II peptide with a His 6 tag.

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]

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 TU Delft iGEM13 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 Magainin II on S. delphini, B. subtilis and E. coli were done according to the protocol described [http://2013.igem.org/Team:TU-Delft/PeptideCharacterization here]. Magainin II did not give a measurable reduction in growth below 40µM (Figure 1), making us decide not to proceed testing, as modeling showed it was not possible to reach these concentrations through expression in E.coli.

Figure 1: MIC of Magainin II on S. delphini

For Other peptides and their characterization please visit [http://2013.igem.org/Team:TU-Delft/PeptideCharacterization Peptide characterization] on TU Delft iGEM13 Wiki

Comparison with two peptides:

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):

Figure 2: MICs of Maximim-H5, Signiferin and Magainin on E. coli

Peptide Production

For more info about the production of the peptide Magainin II, visit [http://2013.igem.org/Team:TU-Delft/Peptides#SUMO Peptide production] on TU Delft iGEM13 Wiki.

Also check the construct used for the peptide production BBa_K1022103 and more characterization.

@@ Line 30: / Line 30: @@
 '''For Other peptides and their characterization please visit [http://2013.igem.org/Team:TU-Delft/PeptideCharacterization Peptide characterization] on TU Delft iGEM13 Wiki'''
+'''Comparison with two peptides:'''
+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):
+[[Image:Coli high conc TUD.jpg|frame|600px|center]]
+Figure 2: MICs of Maximim-H5, Signiferin and Magainin on ''E. coli''
 ==Peptide Production==