Molecular cloning of human adenosine deaminase gene sequences

Using a mixture of synthetic 17-mer oligonucleotides encoding the 64 possible sequences for a peptide of adenosine deaminase as probe, we have isolated a clone for adenosine deaminase mRNA sequences from a collection of T-cell cDNA recombinants. This cDNA clone, phADA-1, contains an insert of 0.8 ki...

Full description

Saved in:
Bibliographic Details
Published in:The Journal of biological chemistry 1983-11, Vol.258 (21), p.12753-12756
Main Authors: Orkin, S H, Daddona, P E, Shewach, D S, Markham, A F, Bruns, G A, Goff, S C, Kelley, W N
Format: Article
Language:English
Subjects:
Adenosine Deaminase - blood
Adenosine Deaminase - genetics
Amino Acid Sequence
Base Sequence
Biological and medical sciences
Biotechnology
Cloning, Molecular
DNA - isolation & purification
DNA - metabolism
Erythrocytes - enzymology
Fundamental and applied biological sciences. Psychology
Genes
Genetic engineering
Genetic technics
Humans
Methods. Procedures. Technologies
Nucleic Acid Hybridization
Nucleoside Deaminases - genetics
Peptide Fragments - analysis
Plasmids
RNA, Messenger - genetics
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Using a mixture of synthetic 17-mer oligonucleotides encoding the 64 possible sequences for a peptide of adenosine deaminase as probe, we have isolated a clone for adenosine deaminase mRNA sequences from a collection of T-cell cDNA recombinants. This cDNA clone, phADA-1, contains an insert of 0.8 kilobase. In addition to the peptide chosen for synthesis of the oligonucleotide probe, the complete DNA sequence predicts 16 other experimentally determined peptides. Mapping of total cellular human DNAs with several restriction enzymes revealed relatively simple patterns of hybridization with phADA-1 as probe, including a polymorphism for PvuII cleavage. In agreement with previous studies, the adenosine deaminase gene was localized by blot hybridization to chromosome 20 in a hybrid cell mapping panel. Using the cDNA as probe, an 18-kilobase EcoRI fragment of human cellular DNA was also cloned in bacteriophage Charon 4A. These adenosine deaminase clones will prove valuable in the full characterization of the cellular gene, molecular analysis of inherited enzyme deficiency associated with immunodeficiency, and regional mapping of human chromosome 20.
ISSN:0021-9258
1083-351X
DOI:10.1016/S0021-9258(17)44027-0