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PMID: 6688808 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Molecular cloning of human adenosine deaminase gene sequences.

The Journal of biological chemistry ·Vol. 258 ·No. 21 ·1983-11-10 ·Pages 12753-6

Orkin SH, Daddona PE, Shewach DS, Markham AF, Bruns GA, Goff SC, Kelley WN

Abstract

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.

MeSH Terms
Adenosine Deaminase/blood,genetics Amino Acid Sequence Base Sequence Cloning, Molecular DNA/isolation & purification,metabolism Erythrocytes/enzymology Genes Humans Nucleic Acid Hybridization Nucleoside Deaminases/genetics Peptide Fragments/analysis Plasmids RNA, Messenger/genetics
Chemicals
Peptide Fragments RNA, Messenger DNA Nucleoside Deaminases Adenosine Deaminase
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Orkin S H
Daddona P E
Shewach D S
Markham A F
Bruns G A
Goff S C
Kelley W N
Article Info
Journal
The Journal of biological chemistry
Abbr.
J Biol Chem
ISSN
0021-9258
Published
1983-11-10
Pages
12753-6
Language
English
Region
United States
NLM ID
2985121R
Subset
IM
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