An efficient strategy for small-scale screening and production of archaeal membrane transport proteins in Escherichia coli

大肠杆菌中古细菌膜转运蛋白小规模筛选和生产的有效策略

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作者:Pikyee Ma, Filipa Varela, Malgorzata Magoch, Ana Rita Silva, Ana Lúcia Rosário, José Brito, Tânia Filipa Oliveira, Przemyslaw Nogly, Miguel Pessanha, Meike Stelter, Arnulf Kletzin, Peter J F Henderson, Margarida Archer

Background

Membrane proteins play a key role in many fundamental cellular processes such as transport of nutrients, sensing of environmental signals and energy transduction, and account for over 50% of all known drug targets. Despite their importance, structural and functional characterisation of membrane proteins still remains a challenge, partially due to the difficulties in recombinant expression and purification. Therefore the need for development of efficient

Significance

Here, we describe an efficient strategy for heterologous production of membrane transport proteins in E. coli. Small-volume cultures (10 mL) produced sufficient amount of proteins to assess their purity and aggregation state. The methods described in this work are simple to implement and can be easily applied to many more membrane proteins.

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