Abstract
The inherited deficiency of the purine salvage enzyme adenosine deaminase is associated with severe B and T-cell dysfunction. In order to investigate the ADA molecule, we have obtained cDNA clones encoding ADA gene sequences. Clones were isolated by metabolic complementation of E. coli Sø200, which has a deletion of the ADA gene. The human T lymphoblast cell line, Molt 4, was the source of mRNA from which cDNA was synthesized. The cDNA sequences were inserted into the Pst 1 site of pBR322 and the recombinant plasmids were used to transform E. coli Sø200. Bacterial colonies that phenotypically expressed adenosine deaminase were isolated by metabolic screening. We obtained cDNA clones that varied in size from 200 to 1000 bp. None of the ADA clones obtained were able to grow on selective media containing the ADA inhibitor deoxycoformycin. This implies that even the shortest clones encode the active site of the enzyme. Restriction mapping and comparison to published sequences showed that all the short clones lie in the central region of the gene. Supported by grant CA26391.
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Danton, M., Coleman, M. ISOLATION OF ADENOSINE DEAMINASE (ADA) cDNA SEQUENCES, CONTAINING THE ACTIVE SITE, BY COMPLEMENTATION IN ESCHERICHIA COLI: 45. Pediatr Res 19, 751 (1985). https://doi.org/10.1203/00006450-198507000-00065
Issue date:
DOI: https://doi.org/10.1203/00006450-198507000-00065
