Yeast can carry out homologous recombination, which is an extremely efficient repair process. Gap-repair cloning (Ma et al., 1987) takes advantage of this process, allowing an insert to be easily cloned into a vector in yeast (Fig. 1).

If you need to make a complicated vector or insert, perform mutagenesis, carry out high-throughput cloning, or otherwise are having trouble with subcloning, we may be able to help. Contact us by email or phone for more details.

For more complicated cloning and mutagenesis, several inserts (with overlapping ends) can be transformed with a gapped yeast vector (Fig. 2). Once recombination has taken place, the DNA insert can be cut out of the yeast vector, and placed into any bacterial or other vector. An advantage of recombination cloning is that the insert does not need to match the restriction enzyme sites on the vector, and in fact, does not need any enzyme sites at all. By using overlapping inserts, proteins or domains can be fused to each other easily, even if they don't have restriction sites.