Beilstein J. Org. Chem.2017,13, 874–882, doi:10.3762/bjoc.13.88
(GFP) fusion protein was functionally expressed in insect Sf9 cells by the BAC-TO-BAC® baculovirusexpressionsystem. A two-step procedure to purify the GAT1/GFP fusion protein from insect Sf9 cells has been established and involves immunoaffinity chromatography using self-prepared anti-GFP antibodies
isolated in its monomeric form.
Keywords: baculovirusexpressionsystem; chromatography; γ-aminobutyric acid (GABA); GABA transporter 1 (GAT1); Introduction
γ-Aminobutyric acid (GABA) is the most abundant inhibitory neurotransmitter in the central nervous system (CNS) of vertebrate species. GABAergic
proteins should be in a native and functional conformation during expression and purification. The natural abundance of most membrane proteins is typically not high enough for the isolation of sufficient quantities for functional and structural studies. In this work, the baculovirusexpressionsystem was
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Graphical Abstract
Figure 1:
Expression of GFP-tagged GAT1 in infected insect cells. (A) Flow cytometry analysis of GAT1/GFP in ...