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High-yield overproduction and purification of human aquaporins from

STAR Protoc. 2022-04; 
Tamim Al-Jubair, Jonas Hyld Steffen, Julie Winkel Missel, Philip Kitchen, Mootaz M Salman, Roslyn M Bill, Pontus Gourdon, Susanna Törnroth-Horsefield
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Gene Synthesis … effect on the expression levels, we strongly recommend to make both N- and C-terminally tagged constructs. … AQP gene with His Tag and TEV cleavage site Synthesis by GenScript N/A … Get A Quote

摘要

Aquaporins (AQPs) are membrane-bound water channels that play crucial roles in maintaining the water homeostasis of the human body. Here, we present a protocol for high-yield recombinant expression of human AQPs in the methylotropic yeast and subsequent AQP purification. The protocol typically yields 1-5 mg AQP per g of yeast cell at >95% purity and is compatible with any membrane protein cloned into , although expression levels may vary. For complete details on the use and execution of this protocol, please refer to Kitchen et al. (2020) and Frick et al. (2014).

关键词

Cell Membrane, Molecular Biology, Protein Biochemistry, Protein expression and purification