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. © 2009 Humana Press.
CITATION STYLE
Berrow, N. S., Alderton, D., & Owens, R. J. (2009). The precise engineering of expression vectors using high-throughput In-FusionTM PCR cloning. Methods in Molecular Biology, 498, 75–90. https://doi.org/10.1007/978-1-59745-196-3_5
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