Homologous Recombination in Mammalian Cells Mediates Formation of a Functional Gene from Two Overlapping Gene Fragments

Chinese hamster cells with a lesion in the CAD gene (cell line Urd-A) require exogenous uridine to survive. Uridine prototrophs could be isolated after introducing two recombinant plasmids containing overlapping fragments of a cloned Syrian hamster CAD gene. In contrast, no uridine prototrophs were...

Full description

Saved in:
Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1983-04, Vol.80 (7), p.2002-2006
Main Authors: Bruno Robert De Saint Vincent, Wahl, Geoffrey M.
Format: Article
Language:English
Subjects:
Animal cells
Animals
Aspartate Carbamoyltransferase - genetics
Carbamoyl-Phosphate Synthase (Glutamine-Hydrolyzing) - genetics
Cell lines
CHO cells
Cricetinae
Crossing Over, Genetic
Dihydroorotase - genetics
DNA
DNA probes
Genes
Genetic vectors
Molecules
Multienzyme Complexes - genetics
Plasmids
Proteins - genetics
Protoplasts
Recombination, Genetic
Sister Chromatid Exchange
Somatic cells
Uridine - biosynthesis
Citations: Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Chinese hamster cells with a lesion in the CAD gene (cell line Urd-A) require exogenous uridine to survive. Uridine prototrophs could be isolated after introducing two recombinant plasmids containing overlapping fragments of a cloned Syrian hamster CAD gene. In contrast, no uridine prototrophs were obtained after introducing a plasmid containing only one of the two overlapping fragments. DNA restriction analysis showed that the prototrophic transformants contain a functional CAD gene which was formed by a recombination event in the overlapping region of the two clones. Most of the recombination events involved homologous exchanges, and some of them apparently were reciprocal. In situ hybridization analysis revealed that the donated sequences were integrated at a single chromosomal site which was different in each transformant. These results demonstrate the existence of a recombination system(s) in mammalian cells that can catalyze homologous exchanges. Recombination between donated sequences is a means by which this system can be characterized and also utilized for the production of novel gene fusions.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.80.7.2002