Evaluation of P. aeruginosa QS Genes

The expression levels of P. aeruginosa QS circuit genes were evaluated by RT-qPCR, as described previously (55 (link)). P. aeruginosa cultures for bacteriophage-resistant strain TL3780-R (TL3780-R1 and TL3780-R2) and the parental strain TL3780 were incubated in fresh LB broth at 37°C under 180 rpm until reaching the logarithmic growth phase (OD₆₀₀ 0.5–0.6). Total RNA was extracted from planktonic bacteria using the RNeasy Mini Kit (Qiagen, Valencia, CA, USA) in accordance with the manufacturer’s instructions. Purified RNA was reverse transcribed onto cDNA using the cDNA Synthesis Kit (TaKaRa, Tokyo, Japan) in accordance with the manufacturer’s instructions. The gene expression levels were measured by qRT-PCR using the TB Green Premix Ex Taq II (Tli RNase H Plus) (2×) (Takara) with specific primers listed in Table S3. rpsL was used as an internal control to normalize the data. The gene expression levels were calculated using the 2^−△△Ct method.