Effect of vector topology on homologous recombination at the CHO aprt locus

The Chinese hamster ovary aprt gene was used as a model for studying the effect of vector topology on gene targeting frequency. A single recombination vector containing 2.7 kb of isogenic DNA homologous to the aprt gene was digested with eight separate restriction enzymes to generate a variety of bo...

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Bibliographic Details
Published inSomatic cell and molecular genetics Vol. 19; no. 3; p. 257
Main Authors Ward, M A, Abramow-Newerly, W, Roder, J C
Format Journal Article
LanguageEnglish
Published United States 01.05.1993
Subjects
Adenine Phosphoribosyltransferase - genetics
Animals
Blotting, Southern
CHO Cells
Cricetinae
Crossing Over, Genetic
Gene Conversion
Genetic Vectors
Recombination, Genetic
Transfection
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Summary:The Chinese hamster ovary aprt gene was used as a model for studying the effect of vector topology on gene targeting frequency. A single recombination vector containing 2.7 kb of isogenic DNA homologous to the aprt gene was digested with eight separate restriction enzymes to generate a variety of both replacement- and insertion-type recombination substrates. The frequency of homologous recombination, normalized by cotransfection with a linearized neo' marker, was assayed by the correction of a mutant hemizygous aprt allele and was not found to reflect vector topology. Southern analysis of representative recombination products suggests that the gene targeting events occurred predominantly by double crossover/gene conversion.
ISSN:0740-7750
DOI:10.1007/BF01233073