Construction of yeast artificial chromosomes containing barley DNA and the identification of clones carrying copies of the repeated element BIS‐1

Abstract

Yeast artificial chromosome (YAC) cloning vectors allow the isolation and analysis of very large segments of DNA. Barley DNA was cleaved with the rare‐cutting restriction enzyme Mlul and fractionated according to size on a linear sucrose gradient or by pulsed field gel electrophoresis. DNA fragments of approximately 50–250 kb were ligated with the YAC vector pYAC‐RC and transformed into yeast spheroplasts. The presence of recombinant YACs with barley DNA inserts was established, and a number of clones containing copies of the repeated element BIS‐1 were isolated. These results show that YAC cloning techniques can be successfully applied to the analysis of the barley genome. © 1991 Mendelian Society of Lund