PCR-based isolation and chromosome assignment of members of the Em gene family from wheat.

Four members belonging to the wheat Em gene family were isolated by PCR, cloned and subsequently sequenced. One of the genes corresponds perfectly to a previous published cDNA sequence, the other three genes are new. The amplified sequences contain the entire coding region, which is interrupted by a short intron of variable length, and part of the 3' untranslated region. The chromosomal assignment of each of the four sequences and three extra, previously published, Em sequences was determined using PCR with sequence-specific primers on wheat aneuploid nullitetrasomic lines. Three sequences were shown to be encoded by the Em-A1 locus (on chromosome 1A), one by Em-B1 on chromosome 1B and two by Em-D1 on chromosome 1D. Hence, primer sets specific for each of the three homoeologous chromosomes of the group 1 are available. A lot of DNA sequence polymorphism exists among the sequences most of which is found in the non-coding parts and mainly in the introns. Sequence alignment groups the seven known Em sequences irrespective of their locus origin. The implication of these findings in relation to the organisation and evolution of the Em gene family are discussed.