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Cloning of terminal transferase cDNA by antibody screening

Landau, Nathaniel R. and St. John, Thomas P. and Weissman, Irving L. and Wolf, Susan C. and Silverstone, Allen E. and Baltimore, David (1984) Cloning of terminal transferase cDNA by antibody screening. Proceedings of the National Academy of Sciences of the United States of America, 81 (18). pp. 5836-5840. ISSN 0027-8424. http://resolver.caltech.edu/CaltechAUTHORS:LANpnas84

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Abstract

A cDNA library was prepared from a terminal deoxynucleotidyltransferase-containing thymoma in the phage vector λgt11. By screening plaques with anti-terminal transferase antibody, positive clones were identified of which some had β-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.


Item Type:Article
Additional Information:© 1984 by the National Academy of Sciences. Contributed by David Baltimore, May 23, 1984. This work was supported by American Cancer Society Grant IM-355 to D.B., National Cancer Institute Grant CA14051 (core grant to S.E. Luria), National Cancer Institute Grant CA22599 and American Cancer Society Grant CH256 to A.E.S., and National Institutes of Health Grant AI18162 to I.L.W. The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.
Subject Keywords:λgt11; pre-B-lymphoid cells
Record Number:CaltechAUTHORS:LANpnas84
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:LANpnas84
Alternative URL:http://dx.doi.org/10.1073/pnas.81.18.5836
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:10040
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:08 Apr 2008
Last Modified:14 Nov 2014 19:20

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