Invention:
Yeast-Agrobacterium-Bacteria (YAB) is a combination of two technologies: yeast recombination capabilities and Agrobacterium tumefaciens-mediated transformation (ATMT). Generation of ATMT constructs usually requires PCR-amplification of DNA fragments and multiple cloning steps into binary Agrobacterium vectors. YAB is a plasmid vector that can be used to combine in one single step several kinds of DNA fragments using the yeast recombination machinery instead of restriction and ligation enzymes, producing a construct that can afterwards be used to introduce the combined DNA fragments into fungi, plants and other suitable organisms, including human cells. University of Arizona faculty created YAB by combining a yeast plasmid with origins of replication for yeast and for bacteria with all the essential parts of an Agrobacterium plasmid necessary to infect cells (plant, fungal) and transfer DNA (Ti plasmid), and origin of replication for Agrobacterium cells. Construction of any plasmid construct designed using the YAB platform can be achieved in one single step.
Background:
Agrobacterium tumefaciens-mediated transformation (ATMT) has become the preferred (and at times the only functional) method to introduce modified genes in many fungal systems. Therefore, there is a need for a simple and efficient method to create gene constructs in an ATMT vector.
Advantages:
- YAB combines the power of yeast recombination in an ATMT-ready vector
- Allows for one-step cloning of multiple inserts
- Can eliminate the need for PCR amplification
- Efficient throughput of constructs for genome analyses of fungi, plants, and other organisms
- YAB constructs introduced in Agrobacterium cells can be used to create genetic modified organisms in fungi, plants, human cells and other suitable organisms without further sub-cloning steps
