Constructing a deletion vector for <i>brpA.</i>

brpA was amplified including 1,000 nucleotides up and down stream of the gene (A) and cloned into the TopoXL vector, constructing Topo-XL::brpA (B). brpA was removed from the vector by PCR amplification adding AvrII and NheI restriction sites (C and D) and the amplicon was double digested. The B. turicatae flgBP-gent was amplified adding AvrII and SpeI restriction sites, double digested, and the knockout vector constructed by ligation (E).