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Increased frequency of N-region insertion in a murine pre-B-cell line infected with a terminal deoxynucleotidyl transferase retroviral expression vector

Landau, Nathaniel R. and Schatz, David G. and Rosa, Margaret and Baltimore, David (1987) Increased frequency of N-region insertion in a murine pre-B-cell line infected with a terminal deoxynucleotidyl transferase retroviral expression vector. Molecular and Cellular Biology, 7 (9). pp. 3237-3243. ISSN 0270-7306. http://resolver.caltech.edu/CaltechAUTHORS:LANmcb87

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Abstract

The role of terminal deoxynucleotidyl transferase (TdT) in the insertion of N regions into the junctional sites of immunoglobulin genes was investigated. Pre-B-cell lines capable of continuous rearrangement of immunoglobulin light-chain genes and differing only in the presence or apparent absence of TdT were derived by infecting cells with a TdT retroviral expression vector or a control vector. The cell lines were then superinfected with a retrovirus-based artificial immunoglobulin gene rearrangement substrate. The substrate was allowed to rearrange in the cell lines and the rearranged proviruses were rescued from the cell lines. Nucleotide sequence analysis of the V-J junctions of the proviral rearranged genes showed a fivefold greater frequency of N-region insertion in proviruses rescued from the TdT+ cell lines than in those rescued from the TdT- cell lines, so that at least 50% of the rearrangements that occurred in the presence of TdT had N regions. It is thus evident that TdT can stimulate N-region insertion, and the enzyme is presumably directly responsible for adding nucleotides at V-J and other immunoglobulin and T-cell receptor gene junctions.


Item Type:Article
Additional Information:Copyright © 1987 by the American Society for Microbiology. Received 4 May 1987/Accepted 5 June 1987. This work was supported by a grant from the American Cancer Society. David G. Schatz is a recipient of a fellowship from the Life and Health Insurance Medical Research Fund.
Record Number:CaltechAUTHORS:LANmcb87
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:LANmcb87
Alternative URL:http://mcb.asm.org/cgi/content/abstract/7/9/3237
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:9984
Collection:CaltechAUTHORS
Deposited By: Archive Administrator
Deposited On:01 Apr 2008
Last Modified:26 Dec 2012 09:55

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