Fig. 1
Southern blotting, and Isolation and sequencing of Xss-V2–18 tal-genes. a Southern blot analysis of BamHI-digested genomic (gDNA) and plasmid DNA (pDNA) of Xcm strain Xss-V2–18. A 2.9-kb SphI fragment of pthXo1 (from Xoo) was labeled with digoxygenin (DIG) and used as a probe to detect tal genes in Xcm Xss-V2–18. b Plasmid DNA of Xss-V2–18 was digested with BamHI, and fragments were gel-purified and ligated into BamHI-digested and CIP-treated pBluescript II SK(−). Southern blot analysis was performed by the using internal SphI fragment of pthXo1 as a probe to confirm each clone (pB-tal1 – pB-tal6). c Schematic diagram of strategy used to sequence tal genes. After cloning into pBluescript II SK(−), the EZ-Tn5™ < KAN-2 > Tnp Transposome™ Kit was used to insert Tn5 into each tal gene. Clones with Tn5 insertions in the middle of the CRR were selected by SphI digestion and sequenced using primer pairs tal-F/RP and FP/tal-R