Journal
STAR PROTOCOLS
Volume 3, Issue 2, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.xpro.2022.101298
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Funding
- Swedish Research Council [2013-05945, 2019-06106]
- Crafoord Foundation [20140711, 20180916]
- Magnus Bergvall Foundation [2015-01534]
- Knut and Alice Wallenberg Foundation [2015.0131, 2020.0194]
- Lundbeck Foundation [R133-A12689, R313-2019-774]
- Independent Research Fund Denmark [4183-00559]
- NordForsk [82000]
- UK Biotechnology & Biosciences Research Council [BB/P025927/1]
- Aston University 50th Anniversary Prize Fellowship
- Biogen
- Swedish Research Council [2019-06106, 2013-05945] Funding Source: Swedish Research Council
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This article presents a protocol for high-yield recombinant expression and purification of human AQPs in the methylotropic yeast Pichia pastoris.
Aquaporins (AQPs) are membrane-bound water channels that play crucial roles in maintaining the water homeostasis of the human body. Here, we present a pro-tocol for high-yield recombinant expression of human AQPs in the methylotropic yeast Pichia pastoris 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 Pichia pastoris, 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).
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