Technical Abstract: The '-gliadins are one of the classes of wheat seed storage proteins, but are the least characterized. In this report, an analysis is made of all available '-gliadin DNA sequences including '-gliadins genes within a large genomic clone, previously reported gene sequences, and ESTs identified from the large wheat EST collection. A contiguous portion of the Gli-B3 locus is shown to contain two apparently active '-gliadin genes, two pseudogenes, and four fragments of the 3' portion of '-gliadin sequences. Results show three groupings of '-gliadin active gene sequences assigned to each of the three hexaploid wheat genomes, and a fourth group thus far consisting of pseudogenes assigned to the A-genome. Analysis of '-gliadin ESTs allows reconstruction of two full-length model sequences encoding the AREL- and ARQL-type proteins from the Gli-A3 and Gli-D3 loci, respectively, and there is no DNA evidence of multiple active genes from these two loci. In contrast, ESTs allow identification of at least 3-4 distinct active genes at the Gli-B3 locus of some cultivars. Additional results from EST and gene analysis include more information on the position of cysteines in some '-gliadin genes and a C-terminal frameshift in one active '-gliadin gene. Problems in studying the '-gliadin gene family are discussed and include high rates of chimeric, frame-shifted, and deleted clones, and cautions interpreting sequences dependent on sequence replications, particularly isolated through PCR.