Genome structure of pTi-SAKURA (I): strategy for DNA sequencing of a Japanese cherry-Ti plasmid.

We isolated a plasmid from a bacterium Agrobacterium tumefaciens, which had been found in a crown gall tumor on a Japanese cherry tree SAKURA and designated it pTi-SAKURA. For complete DNA sequencing, we constructed a DNA library in lambda phage vector and developed a sequencing method by primer walking with long PCR and a PCR subcloning technique for long insert DNA.