A versatile method for efficient YAC transfer between any two strains.
The ability to transfer yeast artificial chromosome (YAC) clones among yeast hosts greatly enhances their utility as cloned DNAs by increasing the range of methods available for experimental manipulation. An effective method for the transfer of YACs between strains in Kar1- matings is described in the accompanying paper (F. Spencer et al., 1994, Genomics 22, 118-126). To evaluate the general nature of the new methodology, we compare YAC transfer in matings in which the YAC donor, the recipient, or both partners carry the kar1 mutation. A set of four universal kar1 intermediary strains that allow YAC transfer from any source to any target strain of the same or of opposite mating type is described. The procedure requires elementary microbial manipulations, including yeast culture and replica plating, and pulsed-field gel electrophoresis for verification of the YAC transfer and integrity. Transfer of YACs by Kar1- mating provides an efficient, reliable, and highly flexible technique that will greatly facilitate YAC manipulation required for a wide variety of applications.
Hugerat
Y
Y
Department of Genetics, Hebrew University, Jerusalem, Israel.
Spencer
F
F
Zenvirth
D
D
Simchen
G
G
D000818:Animals; D018244:Chromosomes, Artificial, Yeast / Q000235:genetics*; D003001:Cloning, Molecular; D018014:Gene Transfer Techniques; D005800:Genes, Fungal; D049770:Genes, Mating Type, Fungal; D005821:Genetic Techniques; D005822:Genetic Vectors; D006801:Humans; D009154:Mutation; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development