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Nucleic Acids Research, 1990, Vol. 18, No. 21 6247-6251
© 1990

MOLECULAR BIOLOGY

Generation of small mutation in large genomic fragments by homologous recombination: description of the technique and examples of its use

Marco Tripodi1, Sabina Perfumo1, Robin Ali1,2, Laura Amicone1, Cathy Abbott2 and Riccardo Cortese3

1Dipartimento di Biopatologia umana, Sezione di Biologia Cellulare Universita' la Sapienza, Policlinico Umberto I Rome, Italy 2MRC Human Biochemical Genetics Unit, University College London 4 Stephenson Way, London NW1 2HE, UK 3European Molecular Biology Laboratory Meyerhofstrasse 1, 6900 Heidelberg, FRG

Received August 2, 1990. Revised September 24, 1990. Accepted September 24, 1990.

We have developed a technique of homologous recombination in bacteria which allows the mutagenesis of large genomic fragments cloned in cosmids. The desired mutation is first introduced into a plasmid clone and is then transferred to the appropriate cosmid clone by the means of double antibiotic selection coupled with phenotypic selection. We describe three different types of construct made by this technique.


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