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PMID: 6091113 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Cloning of terminal transferase cDNA by antibody screening.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 81 ·No. 18 ·1984-09-00 ·Pages 5836-40

Landau NR, St John TP, Weissman IL, Wolf SC, Silverstone AE, Baltimore D

Abstract

A cDNA library was prepared from a terminal deoxynucleotidyltransferase-containing thymoma in the lambda phage vector lambda gt11. By screening plaques with anti-terminal transferase antibody, positive clones were identified of which some had beta-galactosidase-cDNA fusion proteins identifiable after electrophoretic fractionation by immunoblotting with anti-terminal transferase antibody. The predominant class of cross-hybridizing clones was determined to represent cDNA for terminal transferase by showing that one representative clone hybridized to a 2200-nucleotide mRNA in close-matched enzyme-positive but not to enzyme-negative cells and that the cDNA selected a mRNA that translated to give a protein of the size and antigenic characteristics of terminal transferase. Only a small amount of genomic DNA hybridized to the longest available clone, indicating that the sequence is virtually unique in the mouse genome.

MeSH Terms
Animals Antibodies Antigen-Antibody Complex Bacteriophage lambda/genetics Cloning, Molecular DNA/metabolism DNA Nucleotidylexotransferase/analysis,genetics DNA Nucleotidyltransferases/genetics Escherichia coli/genetics Genetic Vectors Mice Nucleic Acid Hybridization Protein Biosynthesis RNA, Messenger/genetics Thymoma/enzymology Thymus Neoplasms/enzymology beta-Galactosidase/genetics
Chemicals
Antibodies Antigen-Antibody Complex RNA, Messenger DNA DNA Nucleotidyltransferases DNA Nucleotidylexotransferase beta-Galactosidase
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Landau N R
St John T P
Weissman I L
Wolf S C
Silverstone A E
Baltimore D
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28 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.81.18.5836
Published
1984-09-00
Pages
5836-40
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC391806
Subset
IM
Grants
NIAID NIH HHS · AI18162 · United States
NCI NIH HHS · CA14051 · United States
NCI NIH HHS · CA22599 · United States
External Links
PubMed source DOI: 10.1073/pnas.81.18.5836 PMC Full Text View journal details
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