Summary
In this chapter, protocols for the construction of expression vectors using In-Fusion™ PCR cloning are presented. The method enables vector and insert DNA sequences to be seamlessly joined in a ligation-independent reaction. This property of the In-Fusion process has been exploited in the design of a suite of multi-host compatible vectors for the expression of proteins with precisely engineered His-tags. Vector preparation, PCR amplification of the sequence to be cloned and the procedure for inserting the PCR product into the vector by In-Fusion™ are described.
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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Berrow, N.S., Alderton, D., Owens, R.J. (2009). The Precise Engineering of Expression Vectors Using High-Throughput In-Fusion™ PCR Cloning. In: Doyle, S.A. (eds) High Throughput Protein Expression and Purification. Methods in Molecular Biology, vol 498. Humana Press. https://doi.org/10.1007/978-1-59745-196-3_5
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DOI: https://doi.org/10.1007/978-1-59745-196-3_5
Publisher Name: Humana Press
Print ISBN: 978-1-58829-879-9
Online ISBN: 978-1-59745-196-3
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