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 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.
Ward
M A
MA
Department of Molecular and Medical Genetics, University of Toronto, Samuel Lununfeld Research Institute, Mount Sinai Hospital, Ontario, Canada.
Abramow-Newerly
W
W
Roder
J C
JC
D000228:Adenine Phosphoribosyltransferase / Q000235:genetics*; D000818:Animals; D015139:Blotting, Southern; D016466:CHO Cells; D006224:Cricetinae; D003434:Crossing Over, Genetic; D005785:Gene Conversion; D005822:Genetic Vectors; D011995:Recombination, Genetic; D014162:Transfection
D000228:Adenine Phosphoribosyltransferase