In-Fusion™ assembly: seamless engineering of multidomain fusion proteins, modular vectors, and mutations

In-Fusion™ can join any two pieces of DNA that have a 15-bp overlap at their ends. The result is equivalent to a recombination event at the ends of the DNAs. The 15-bp overlap may be engineered by inclusion in primers used to PCR amplify a segment of DNA. Originally described for inserting one piece...

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Bibliographic Details
Main Authors: Baogong Zhu, Guifang Cai, Emily O. Hall, Gordon J. Freeman
Format: Article
Language:English
Published: Taylor & Francis Group 2007-09-01
Series:BioTechniques
Online Access:https://www.future-science.com/doi/10.2144/000112536
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Summary:In-Fusion™ can join any two pieces of DNA that have a 15-bp overlap at their ends. The result is equivalent to a recombination event at the ends of the DNAs. The 15-bp overlap may be engineered by inclusion in primers used to PCR amplify a segment of DNA. Originally described for inserting one piece of DNA into a restriction enzyme-digested plasmid, We have found In-Fusion can join four or more pieces of DNA in a single reaction. We used this insight to construct seamless fusion proteins, modular vectors with readily interchangeable segments, and novel mutagenesis strategies. Replacement In-Fusion can be used to delete any desired DNA segment in a plasmid and replace it with any desired new DNA segment without limitations on position or size.
ISSN:0736-6205
1940-9818