Article
Markerless gene replacement in Escherichia coli stimulated by a double-strand break in the chromosome.
Nucleic acids research - 15 Nov 1999
Pósfai G, Kolisnychenko V, Bereczki Z, Blattner F R
Abstract excerpt
A simple and efficient gene replacement method, based on the recombination and repair activities of the cell, was developed. The method permits the targeted construction of markerless deletions, insertions and point mutations in the Escherichia coli chromosome. A suicide plasmid, carrying the mutant allele and the recognition site of meganuclease I- Sce I, is inserted into the genome by homologous recombination...
Topics
- Chromosome Deletion
- Chromosomes, Bacterial
- DNA
- DNA, Bacterial
- Deoxyribonucleases, Type II Site-Specific
- Electroporation
- Escherichia coli
- Gene Transfer Techniques
- Molecular Sequence Data
- Mutation
- Recombination, Genetic
- Saccharomyces cerevisiae Proteins