Part:BBa_K5248055
pACYCDuet-1-mCherry-mchI（6his)-EGFP-mchB(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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E. coli BL21 DE3 containing the expression plasmid of the mchB gene, (ii) LB blank validation, (iii) Fusobacterium bezoarum ADP1 without the expression plasmid of the mchB gene, (iv) E. coli BL21 without the expression plasmid of the mchB gene, and (v) E. coli BL21 DE3 without the expression plasmid of the mchB 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 143
Illegal PstI site found at 369
Illegal PstI site found at 378 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 143
Illegal NheI site found at 589
Illegal PstI site found at 369
Illegal PstI site found at 378 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 143
- 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 143
Illegal PstI site found at 369
Illegal PstI site found at 378 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 143
Illegal PstI site found at 369
Illegal PstI site found at 378 - 1000COMPATIBLE WITH RFC[1000]
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