Development of an experimental method of systematically estimating protein expression limits in HEK293 cells.

ABSTRACT: Protein overexpression sometimes causes cellular defects, although the underlying mechanism is still unknown. A protein's expression limit, which triggers cellular defects, is a useful indication of the underlying mechanism. In this study, we developed an experimental method of estimating the expression limits of target proteins in the human embryonic kidney cell line HEK293 by measuring the proteins' expression levels in cells that survived after the high-copy introduction of plasmid DNA by which the proteins were expressed under a strong cytomegalovirus promoter. The expression limits of nonfluorescent target proteins were indirectly estimated by measuring the levels of green fluorescent protein (GFP) connected to the target proteins with the self-cleaving sequence P2A. The expression li

SUBMITTER: Mori Y 

PROVIDER: S-EPMC7075890 | biostudies-literature | 2020 Mar

REPOSITORIES: biostudies-literature