Screening of Membrane Protein Production by Comparison of Transient Expression in Insect and Mammalian Cells.

ABSTRACT: Membrane proteins are difficult biomolecules to express and purify. In this paper, we compare the small-scale production of six selected eukaryotic integral membrane proteins in insect and mammalian cell expression systems using different techniques for gene delivery. The target proteins were C terminally fused to the green fluorescent marker protein GFP to enable sensitive monitoring. We show that the choice of expression systems makes a considerable difference to the yield and quality of the six selected membrane proteins. Virus-free transient gene expression (TGE) in insect High Five cells combined with solubilization in dodecylmaltoside plus cholesteryl hemisuccinate generated the most homogeneous samples for all six targets. Further, the affinity purification of the solubilized protei

SUBMITTER: Kaipa JM 

PROVIDER: S-EPMC10216445 | biostudies-literature | 2023 May

REPOSITORIES: biostudies-literature