Establishment of mammalian cell lines containing multiple nonsense mutations and functional suppressor tRNA genes.

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Publication Type Journal Article
School or College College of Science; School of Medicine
Department Oncological Sciences; Biology; Human Genetics
Program Institute of Human Genetics; Howard Hughes Medical Institute (HHMI)
Creator Capecchi, Mario R.
Other Author Hudziak, Robert M.; Laski, Frank A.; RajBhandary, Uttam L.; Sharp, Phillip A.
Title Establishment of mammalian cell lines containing multiple nonsense mutations and functional suppressor tRNA genes.
Date 1982-11-30
Description We describe the generation of mammalian cell lines carrying amber suppressor genes. Nonsense mutants in the herpes simplex virus thymidine kinase (HSV tk) gene, the Escherichia coli xanthine-guanine phosphoribosyl transferase (Eco-gpt) gene and the aminoglycoside 3' phosphotransferase gene of the Tn5 transposon (NPT-II) were isolated and characterized. Each gene was engineered with the appropriate control signals to allow expression in both E. coli and mammalian cells. Expression in E. coli made possible the use of well developed bacterial and phage genetic manipulations to isolate and characterize the nonsense mutants. Once characterized, the nonsense mutants were transferred into mammalian cells by microinjection and used, in turn, to select for amber suppressor genes. Xenopus laevis amber suppressor genes, prepared by site-specific mutagenesis of a normal X. laevis tRNA gene, were microinjected into the above cell lines and selected for the expression of one or more of the amber mutant gene products. The resulting cell lines, containing functional amber suppressor genes, are stable and exhibit normal growth rates.
Publisher MIT
Volume 31
Issue 1
First Page 137
Last Page 146
Subject Cercopithecus aethiops; Escherichia coli; Xenopus
Subject MESH Mutation; RNA, Transfer; Suppression, Genetic
Language eng
Bibliographic Citation Cell. 1982 Nov;31(1):137-46: Hudziak RM, Laski FA, RajBhandary UL, Sharp PA, Capecchi MR. Establishment of mammalian cell lines containing multiple nonsense mutations and functional suppressor tRNA genes. Retrieved on September 22.2006 from http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6WSN-4C8973V-20&_coverDate=11%2F30%2F1982&_alid=454357817&_rdoc=1&_fmt=&_orig=search&_qd=1&_cdi=7051&_sort=d&view=c&_acct=C000049956&_version=1&_urlVersion=0&_userid=990933&md5=1854d3b8ba32689d8abb128fab25d12e.
Rights Management Copyright © 1982, MIT. All Rights Reserved.
Format Medium application/pdf
Identifier ir-main,467
ARK ark:/87278/s69k4vsb
Setname ir_uspace
Date Created 2012-06-13
Date Modified 2012-06-13
ID 706235
Reference URL https://collections.lib.utah.edu/ark:/87278/s69k4vsb
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