Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
4
pubmed:dateCreated
1985-6-13
pubmed:databankReference
pubmed:abstractText
Human adenosine deaminase (ADA) is an important purine catabolic enzyme which irreversibly deaminates adenosine and deoxyadenosine. Severe genetic deficiency of ADA leads to an immunological deficiency state in which T-lymphoid cells are selectively destroyed by the accumulation of toxic levels of deoxyadenosine and deoxy-ATP. In preparation for transfer of ADA sequences into a variety of cell types, we explored expression of ADA cDNAs transfected into cultured cells within a simian virus 40-based expression vector. After transfection into monkey kidney (COS) cells, ADA cDNA encompassing the entire coding region of the protein generated human ADA activity. An unexpected finding, however, was the identification of a cDNA clone that failed to produce either human enzyme activity or immunoreactive ADA protein. As this pattern is typical of many naturally occurring mutant ADA alleles, we characterized the molecular defect in this clone. DNA sequence analysis revealed a single nucleotide substitution in amino acid position 50 (glycine-valine). Northern blotting with a unique 17-mer oligonucleotide demonstrated the absence of the mutant sequence in the mRNA from which the cDNA library giving rise to the mutant cDNA was constructed. Therefore, the substitution in the variant cDNA was created during cloning. These data define one critical region of the human ADA protein molecule and suggest a convenient strategy for characterization of the phenotypes associated with naturally occurring mutant alleles.
pubmed:grant
pubmed:commentsCorrections
http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-1089883, http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-13062, http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-14907713, http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-215915, http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-372236, http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-4117384, http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-5432063, http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-6090454, http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-6091052, http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-6134754, http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-6146135, http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-6154876, http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-6188062, http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-6198240, http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-6198631, http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-6200875, http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-6208479, http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-6246368, http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-6251420, http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-6255772, http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-6260373, http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-6267992, http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-6312838, http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-6322609, http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-6546794, http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-6603477, http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-6688808, http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-6694735, http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-6744418, http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-6777776, http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-6826735, http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-6863546, http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-6977542, http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-7072935, http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-7177196, http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-7380831, http://linkedlifedata.com/resource/pubmed/commentcorrection/3838797-9388
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Apr
pubmed:issn
0270-7306
pubmed:author
pubmed:issnType
Print
pubmed:volume
5
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
762-7
pubmed:dateRevised
2009-11-18
pubmed:meshHeading
pubmed:year
1985
pubmed:articleTitle
Transient expression of human adenosine deaminase cDNAs: identification of a nonfunctional clone resulting from a single amino acid substitution.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't