Abstract
In vivo recombinational cloning in yeast is the efficient method for the cloning of multiple DNA fragments and large DNA fragments exceeding 10 kbp. Still, this cloning method has been limited to experiments with yeast vectors because many cloning vectors lack yeast replication origins. Therefore, we developed a new system to apply the yeast-recombinational cloning to T-DNA binary vectors, such as pCAMBIA. A newly constructed helper plasmid pSU7 and pSU23 allows the in vivo conversion of plant vectors into vectors replicable in yeast. The DNA fragments to be cloned are PCR-amplified with the addition of homologous sequences to a vector. Cotransformation of the helper plasmid into yeast with the vector and PCR-amplified DNA fragments, results in the conversion of the vector into a vector replicable in yeast and simultaneously allows cloning of the DNA fragments into the same vector.
