Application of modified differential display technique for cloning and sequencing of the 3' region from three putative members of wheat HSP70 gene family

We have modified the differential display technique to isolate 3' regions from different members of the wheat HSP70 gene family. An HSP70 gene family-specific degenerate primer was used as a 5' primer in place of the arbitrary primer used in the original technique. We cloned and sequenced three cDNA fragments that were differentially expressed in heat stressed wheat seedlings. Based on the high similarity between predicted translation products of these three sequences and known members of the HSP70 family from plants, these cDNAs were identified as members of the HSP70 gene family. Two of these members appeared distinct in the 3' non-coding region with only 48% identity. Therefore differential display could successfully be used to isolate 3' regions of different members of a multigene family in a relatively short period, even if the members had highly similar protein-coding regions.