Genetic engineering using homologous recombination

DL Court, JA Sawitzke… - Annual review of genetics, 2002 - annualreviews.org
DL Court, JA Sawitzke, LC Thomason
Annual review of genetics, 2002annualreviews.org
▪ Abstract In the past few years, in vivo technologies have emerged that, due to their
efficiency and simplicity, may one day replace standard genetic engineering techniques.
Constructs can be made on plasmids or directly on the Escherichia coli chromosome from
PCR products or synthetic oligonucleotides by homologous recombination. This is possible
because bacteriophage-encoded recombination functions efficiently recombine sequences
with homologies as short as 35 to 50 base pairs. This technology, termed recombineering, is …
Abstract
In the past few years, in vivo technologies have emerged that, due to their efficiency and simplicity, may one day replace standard genetic engineering techniques. Constructs can be made on plasmids or directly on the Escherichia coli chromosome from PCR products or synthetic oligonucleotides by homologous recombination. This is possible because bacteriophage-encoded recombination functions efficiently recombine sequences with homologies as short as 35 to 50 base pairs. This technology, termed recombineering, is providing new ways to modify genes and segments of the chromosome. This review describes not only recombineering and its applications, but also summarizes homologous recombination in E. coli and early uses of homologous recombination to modify the bacterial chromosome. Finally, based on the premise that phage-mediated recombination functions act at replication forks, specific molecular models are proposed.
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