Expression Screening of Human Integral Membrane Proteins Using BacMam.

Pravin Mahajan, Katherine Ellis,Shubhashish Mukhopadhyay,Alejandra Fernandez-Cid,Gamma Chi,Henry Man,Katharina L Dürr,Nicola A Burgess-Brown

STRUCTURAL GENOMICS, 2 EDITION: General Applications（2021）

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摘要

This chapter describes the step-by-step methods employed by the Structural Genomics Consortium (SGC) for screening and producing proteins in the BacMam system. This eukaryotic expression system was selected and a screening process established in 2016 to enable production of highly challenging human integral membrane proteins (IMPs), which are a significant component of our target list. Here, we discuss our recently developed platform for identifying expression and monodispersity of IMPs from 3 mL of HEK293 cells.

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关键词

BacMam,Baculovirus,FSEC,Gel filtration,IMAC,Insect cells,Mammalian cells,Protein purification,SEC chromatography,Transduction

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