Identification of a virulence tal gene in the cotton pathogen Xanthomonas citri pv. malvacearum strain Xss-V2–18

摘要

Southern blotting, and Isolation and sequencing of Xss-V –18 genes. Southern blot analysis of HI-digested genomic (gDNA) and plasmid DNA (pDNA) of strain Xss-V –18. A 2.9-kb I fragment of (from ) was labeled with digoxygenin (DIG) and used as a probe to detect genes in Xss-V –18. Plasmid DNA of Xss-V –18 was digested with HI, and fragments were gel-purified and ligated into HI-digested and CIP-treated pBluescript II SK(−). Southern blot analysis was performed by the using internal I fragment of as a probe to confirm each clone (pB- – pB- ). Schematic diagram of strategy used to sequence genes. After cloning into pBluescript II SK(−), the EZ-Tn5™  Tnp Transposome™ Kit was used to insert Tn into each gene. Clones with Tn insertions in the middle of the CRR were selected by I digestion and sequenced using primer pairs tal-F/RP and FP/tal-R