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Le projet „Génome Humain” et la caractérisation des étiquettes (E.S.T.) de testicule humain
Sequencing of the human genome and characterization of expressed sequence Tags (E.S.T.) in human testis
Andrologie volume 9, pages 342–346 (1999)
Résumé
L'une des approches développées dans le cadre du projet „Génome Humain”, consiste à caractériser les ARNm d'un tissu donné par la séquence partielle des ADNc correspondants. Les séquences obtenues, ou „Expressed Sequence Tag” (E.S.T.), constituent une source d'information majeure, pour l'identification de nouveaux gènes et leur localisation chromosomique. A ce jour 1 200 000 E.S.T. ont été déposés dans la base de données „dbest” par la communauté scientifique, dont 50 000 ont été obtenus à partir de testicule humain. A partir de ces E.S.T., nous avons caractérisé de nouveaux gènes exprimés spécifiquement dans le testicule humain et notamment une nouvelle mono ADP ribosyl transférase ainsi que deux protéines liant les queues polyA des ARNm.
Abstract
The “Human Genome” is devoted to the identification of all the human genes and their chromosomal localization. Besides the sequencing of the genome, complementary strategies have been developed, including the characterization of mRNA by single pass sequencing of the corresponding cDNA. The partial sequences generated during this process represent expressed sequence tags (E.S.T.) of the corresponding genes. We used this strategy, associated with Northern blot andin situ hybridization, in order to characterize genes expressed in human testis. A cDNA library was prepared using mRNA purified from testes of a 27 years old man. We isolated, and stored in glycerol 7750 recombinant clones. 2200 clones with an insert over 700bp were single-pass sequenced, mainly at the 5' end, generating 2563 sequences products which were compared, with the sequences present in public databases. Out of 2000 sequences 481 were found identical to known human sequences; 358 were homologous to sequences from human or other species; 719 represented new sequences; 442 were rejected. Some of the clones corresponding to mRNAs transcribed from new genes were further analyzed on Northern blot of human, rat and mouse testes mRNA along with RNA of kidney, liver and brain. This allowed us to identify several new genes expressed in human testis of potential interest for the physiology of this tissue. Among others, they include a new mono-ADP ribosyl transferase and two testis specific poly A binding proteins.
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Guellaën, G. Le projet „Génome Humain” et la caractérisation des étiquettes (E.S.T.) de testicule humain. Androl. 9, 342–346 (1999). https://doi.org/10.1007/BF03034807
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DOI: https://doi.org/10.1007/BF03034807