Here, we provide the protocols for the various experiments involved in our project for future use by other teams.
Molecular cloning was applied to the construction of all plasmids in our project.
We verified the expression and roughly determined the content of AHLs degrading enzymes and their mutants, glycosidases degrading biological periplasm in our project by protein induced expression and purification, in order to initially select the more efficient enzymes for the next validation experiments.
We designed multiple characterization experiments by observing the results of enzyme action to verify that the exogenous proteins expressed in our project functioned as expected, and selected superior proteins to construct our SRBioQuencher.
In order to better design the suicide system, we determined the effect of our regulatory gene cluster on the growth of E. coli and verified the sensitivity and preference of our regulatory system to AHLs.
We construct engineering bacteria that secrete antimicrobial peptides and culture SRB for antibacterial experimental verification.