Current Protocols in Molecular Biology · 2007 · 635 citations · 6 references
Life SciencesGeneticsMicrobiologyBacteriologyGenetic EngineeringRecombinationBacteriophage GeneticsBacteriophage VectorsGenetic Engineering Of MicroorganismsHorizontal Gene TransferMolecular CloningPhage GeneticsBacterial Genome EngineeringBacteriophage BiologyBacteriophage-mediated TransferEscherichia ColiPhage Gene TransferPhage TransductionRecombination ReactionsRecombination-based Genetic Engineering
P1 bacteriophage can transfer ~100 kb DNA fragments. This unit outlines a method to transfer segments of the E. coli genome between strains. The method uses a P1 lysate from a donor strain to infect a recipient, where recipient enzymes recombine donor DNA fragments into the recipient chromosome.
Abstract This unit describes the procedure used to move portions of the E. coli genome from one genetic variant to another. Fragments of ∼100 kb can be transferred by the P1 bacteriophage. The phage is first grown on a strain containing the elements to be moved, and the resulting phage lysate is used to infect a second recipient strain. The lysate will contain bacterial DNA as well as phage DNA, and genetic recombination, catalyzed by enzymes of the recipient strain, will incorporate the bacterial fragments into the recipient chromosome. Curr. Protoc. Mol. Biol . 79:1.17.1‐1.17.8. © 2007 by John Wiley & Sons, Inc.
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