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Integrases are essential for T3SS and CRISPR island excision. The two-stage attB PCR amplification assay was used to detect excision of VPI-3 in ΔintV2 and wild-type NRT36S (A) and of VPI-6 in ΔintV1 and wild-type RC385 (B). No PCR product was detected in the first round of PCR and wild-type attB excision products of 524 bp and 210 bp were detected in the second round of PCR for NRT36S and RC385, respectively, and no product was detected for NRT36S ΔintV2 or RC385 ΔintV1.
