Topic Introduction

Gateway Recombinational Cloning

  1. Albertha J.M. Walhout

Abstract

The Gateway recombinatorial cloning system was developed for cloning multiple DNA fragments in parallel (e.g., in 96-well formats) in a standardized manner using the same enzymes. Gateway cloning is based on the highly specific integration and excision reactions of bacteriophage λ into and out of the Escherichia coli genome. Because the sites of recombination (“att” sites) are much longer (25–242 bp) than restriction sites, they are extremely unlikely to occur by chance in DNA fragments. Therefore, the same recombination enzyme can be used to robustly clone many different fragments of variable size in parallel reactions.

Footnotes

  • From the Molecular Cloning collection, edited by Michael R. Green and Joseph Sambrook.

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  1. doi:10.1101/pdb.top094912 Cold Spring Harb Protoc 2018: pdb.top094912- © 2018 Cold Spring Harbor Laboratory Press
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