In an experiment designed to find sequences common to a skeletal muscle cDNA library and an X chromosome specific library, we have isolated cDNA clones corresponding to glyceraldehyde 3 phosphate dehydrogenase (GAPD), (whose gene is assigned to chromosome 12), and a DNA fragment from the X chromosome short arm which contains an intron‐less GAPD pseudogene. A 1210‐bp cDNA sequence has been established which covers all of the protein‐coding region, most of the 5′ non‐coding region and part of the 3′ non‐coding region. It corresponds to the major (and possibly unique) GAPD mRNA present in skeletal muscle. Unexpectedly, the amino acid sequence derived from the cDNA clones differs at 10% of the residues from that established for the human protein purified from skeletal muscle. The X‐linked pseudogene has been localised in the p22‐p11 region of the human X chromosome. It has the structure of a complete retrotranscript of a processed mRNA, including the poly(A) tail and is 96% homologous to the cDNA sequence. The pseudogene is flanked by a 15‐bp direct repeat, and an Alu‐like sequence is found in the 3′‐flanking region. About 25 GAPD sequences are found in the human genome, 12 of which have high homology to the cDNA probe. A similar complexity is found in hamster. In contrast, the mouse genome contains an amazing number of GAPD related fragments (at least 200). The hybridization pattern suggests that this multiplicity has been generated by two different mechanisms: first the generation of approximately 40 different sequences, which were subsequently amplified (probably by tandem duplication).

Dystrophin is a very large muscle protein (approximately 400 kd) the deficiency of which is responsible for Duchenne muscular dystrophy. Its function is unknown at present. In order to know whether different domains of the protein are differentially conserved during evolution, we have cloned and sequenced the chicken dystrophin cDNA. The protein coding sequence has almost the same size as in man. The N‐terminal region that resembles the actin binding domain of alpha actinin, as well as the large spectrin like domain show 80% and 75% conservation respectively between chicken and man. In contrast, the C‐terminal region shows 95% identity over 627 aa suggesting that it is an important region of interaction with other proteins. Comparison of the amino acid sequence of this C‐terminal region to other protein sequences shows only marginally significant similarities. Finally we have found a striking conservation of three segments of the 3′ untranslated sequence (85% homology over a total of 920 nt) between chicken and man. These also appear to be conserved in other mammals. This high conservation is not linked to open reading frames.

To analyse the relationship between DNA undermethylation at some sites in the ovalbumin and conalbumin gene regions (1) and the expression of these genes in chick oviduct, digestions with HhaI, which differentiates between methylated and unmethylated HhaI restriction sites, was performed on DNA isolated from chicken erythrocyte or oviduct chromatin treated with DNase I which degrades preferentially "active" chromatin. This was followed by analysis with ovalbumin- and conalbumin-specific hybridization probes. We conclude that the residual DNA methylation found at some sites of the ovalbumin and conalbumin gene regions is derived from the fraction of cells in which the chromatin of these genes is not in an "active" form. On the other hand, the ovalbumin and conalbumin sites which are partially unmethylated in erythrocyte DNA correspond to chromatin regions which are not DNase I-senitive. We have also detected a site about 1 kb downstream from the 3' end of the conalbumin gene that is hypersensitive to DNase I in all tissues tested.