The amphibian Xenopus laevis has been a central model for the study of evolution of the major histocompatibility complex (MHC). Many of the counterparts of mammalian MHC genes have been identified in Xenopus, facilitating the understanding of MHC structure and function. Herein we characterize X. laevis MHC class II-alpha chain genes. There are three related class II A genes/haplotype in the four commonly used partially inbred strains, all of which linked to the functional MHC. At least two of these genes in the f haplotype encode full-length cDNA clones and a genomic fragment encoding the immunoglobulin-like domain of the third gene was also characterized. The protein structure and domain organization deduced from the two f/f cDNA clones are similar to mammalian MHC class II-alpha chains. Expression of class II A genes is highest in the spleen and intestine, similar to the previously examined tissue distribution of class II B genes. The two highly expressed genes display high sequence diversity among alleles, similar to what has been found in most other species. Surprisingly, transcript sizes of class II A alleles/isotypes are diverse, suggesting that Xenopus class II allelic lineages are very old.