Mapping of the Hor2 locus in barley by pulsed field gel electrophoresis.

Abstract

High molecular weight DNA released from isolated protoplasts was digested with rare-cutting restriction enzymes and separated by pulsed field gel electrophoresis. The average size of undigested DNA was above 1500 kbp. Digests made with NotI, SfiL, Mlul and SalI was hybridized to a probe, common to all genes of the Hor2 locus encoding B-hordein polypeptides, and this revealed the maximum size of the locus to be 360 kbp. Two probes, specific for individual B-hordein genes, enabled the identification of two fragment classes in the locus, each containing an equal number of B-hordein genes. Double digests allowed ordering of sites and construction of a map covering 650 kbp around the Hor2 locus. No evidence for physical linkage of the two fragment classes was obtained. The possible assignment of the two classes of hybridizing fragments to the B1- and B3-hordein subgroups is discussed.