Part:BBa_K5248058
pETduet-1-mcmI-2（6his）-mcmA(strep)
We inoculated 100 μL of each predator strain's overnight concentrated culture on LB agar plates, covered with soft LB agar containing prey strains. For the mcmA and mchB antimicrobial peptide expression plasmids, the prey strain was Escherichia coli DH5α, and the predator strains were E. coli Nissle 1917 DE3 and BL21 DE3. In this experiment, the first two plates showed predators expressing mcmA and mchB, with the prey sensitive to these antimicrobial peptides.
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(i)E. coli 1917 DE3 containing an expression plasmid for the mcmA gene, (ii) LB blank validation, (iii) Fusobacterium bezoarum ADP1 without an expression plasmid for the mcmA gene, (iv) E. coli 1917 without an expression plasmid for the mcmA gene, and (v) E. coli 1917 DE3 without an expression plasmid for the mcmA gene.
After overnight incubation, we observed that the strains containing our designed mcmA or mchB expression plasmids showed visible zones of inhibition. In addition, we verified the growth of these two antimicrobial peptide expression plasmid strains.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 166
- 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 166
- 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 166
Illegal BamHI site found at 559 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 166
- 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 166
- 1000COMPATIBLE WITH RFC[1000]
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