NDLI: Toward an animal model of cystic fibrosis: targeted interruption of exon 10 of the cystic fibrosis transmembrane regulator gene in embryonic stem cells.

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Toward an animal model of cystic fibrosis: targeted interruption of exon 10 of the cystic fibrosis transmembrane regulator gene in embryonic stem cells.

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Toward an animal model of cystic fibrosis: targeted interruption of exon 10 of the cystic fibrosis transmembrane regulator gene in embryonic stem cells.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Boucher, R. C. Latour, A. M. Smithies, O. Wainwright, B. Brigman, K. Scambler, P. Kim, H. S. Koller, B. H.
Description	Author Affiliation: Koller BH ( Department of Pathology, University of North Carolina, Chapel Hill 27599-7525.);
Abstract	A gene-targeting construct was made containing 7.8 kilobases of DNA spanning exon 10 of the mouse cystic fibrosis transmembrane regulator (CFTR) gene in which part of the exon has been replaced by two neomycin-resistance (Neo) genes driven by different promoters. (This replacement introduces a chain-termination codon at amino acid position 489 in the CFTR sequence). A herpes simplex thymidine kinase gene was on each end of the construct, which was electroporated into embryonic stem (ES) cells. Colonies resistant to G418, or to G418 plus ganciclovir, were selected and screened by Southern blotting or by PCR amplification. Five pools of G418-resistant cells gave PCR products diagnostic of targeting. Four independent clones of ES cells with a disrupted CFTR gene have been isolated from these pools. The frequency of targeting was 1/2500 G418-resistant colonies. This low frequency is not the consequence of marginal expression of the Neo genes in the targeted cells. The CFTR targeting events were clustered among our experiments in a manner suggesting that some unidentified factor(s), possibly passage number, influences the recovery of CFTR-targeted cells.
ISSN	00278424
e-ISSN	10916490
Journal	Proceedings of the National Academy of Sciences of the United States of America
Issue Number	23
Volume Number	88
Language	English
Publisher	National Academy of Sciences
Publisher Date	1991-01-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Cystic Fibrosis Genetics Exons Membrane Proteins Stem Cells Physiology Amino Acid Sequence Animals Blotting, Southern Cells, Cultured Codon Cystic Fibrosis Transmembrane Conductance Regulator DNA Disease Models, Animal Drug Resistance, Microbial Embryo, Mammalian Genetic Techniques Kanamycin Kinase Mice Molecular Sequence Data Neomycin Pharmacology Oligodeoxyribonucleotides Phosphotransferases Plasmids Polymerase Chain Reaction Promoter Regions, Genetic Restriction Mapping Simplexvirus Thymidine Kinase Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Multidisciplinary
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

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