Linked Life Data

2410423

Source:http://linkedlifedata.com/resource/pubmed/id/2410423

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
18
pubmed:dateCreated
1985-9-25
pubmed:databankReference
pubmed:abstractText
Total poly(A+) RNA derived from a mouse cell line with amplified adenosine deaminase genes was used as template to synthesize double-stranded cDNA. The cDNAs were inserted into the PstI site of the beta-lactamase gene in plasmid pBR322 following G-C tailing. After transformation into adenosine deaminase-deficient Escherichia coli hosts, recombinant plasmids containing functional murine adenosine deaminase cDNAs were identified by selecting for functional complementation. Analysis of plasmids containing functional adenosine deaminase cDNA sequences strongly suggested that adenosine deaminase expression resulted mainly from beta-lactamase/adenosine deaminase fusion proteins even when the adenosine deaminase codons were out-of-frame with respect to the beta-lactamase gene codons upstream. The nucleotide sequence of a 1.65-kilobase pair cDNA insert in one of the functional recombinant clones was determined and found to contain a 1056-nucleotide open reading frame. When this 1056-nucleotide open reading frame was inserted into a mammalian expression vector and introduced into monkey kidney cells, a high level of authentic mouse adenosine deaminase was produced. Nucleic acid blot analysis using a full-length adenosine deaminase cDNA clone as probe revealed that the mouse adenosine deaminase structural gene was at least 21 kilobase pairs in size and encoded three polyadenylated mRNAs. Analysis of the cDNA library from which the functional clones were isolated suggested that this approach of cloning functional mammalian adenosine deaminase cDNA clones by genetic complementation of enzyme-deficient bacteria could be accomplished even if the abundance of the adenosine deaminase mRNA sequences were as low as approximately 0.001%.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Aug
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
25
pubmed:volume
260
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
10299-307
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed-meshheading:2410423-Adenosine Deaminase, pubmed-meshheading:2410423-Amino Acid Sequence, pubmed-meshheading:2410423-Animals, pubmed-meshheading:2410423-Base Sequence, pubmed-meshheading:2410423-Cell Line, pubmed-meshheading:2410423-Cercopithecus aethiops, pubmed-meshheading:2410423-Cloning, Molecular, pubmed-meshheading:2410423-DNA, pubmed-meshheading:2410423-DNA Transposable Elements, pubmed-meshheading:2410423-Escherichia coli, pubmed-meshheading:2410423-Gene Amplification, pubmed-meshheading:2410423-Genes, pubmed-meshheading:2410423-Genetic Complementation Test, pubmed-meshheading:2410423-Genetic Vectors, pubmed-meshheading:2410423-Kidney, pubmed-meshheading:2410423-Mice, pubmed-meshheading:2410423-Nucleic Acid Hybridization, pubmed-meshheading:2410423-Nucleoside Deaminases, pubmed-meshheading:2410423-Plasmids, pubmed-meshheading:2410423-Poly A, pubmed-meshheading:2410423-RNA, pubmed-meshheading:2410423-RNA, Messenger, pubmed-meshheading:2410423-Thymidine Kinase
pubmed:year
1985
pubmed:articleTitle
Identification of functional murine adenosine deaminase cDNA clones by complementation in Escherichia coli.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, U.S. Gov't, Non-P.H.S., Research Support, Non-U.S. Gov't