Production of a urokinase plasminogen activator-IgG fusion protein (uPA-IgG) in the baculovirus expression system

Gene ◽  
1997 ◽  
Vol 190 (1) ◽  
pp. 139-144 ◽  
Author(s):  
Thomas A. Kost ◽  
Diane M. Ignar ◽  
William C. Clay ◽  
John Andrews ◽  
Jeremy D. Leray ◽  
...  
Keyword(s):  
Fusion Protein ◽  
Plasminogen Activator ◽  
Expression System ◽  
Baculovirus Expression System ◽  
Urokinase Plasminogen Activator ◽  
Baculovirus Expression

scholarly journals Expression, purification and structural analysis of functional GABA transporter 1 using the baculovirus expression system

2017 ◽  
Vol 13 ◽  
pp. 874-882 ◽  
Author(s):  
Jing Hu ◽  
Chris Weise ◽  
Christoph Böttcher ◽  
Hua Fan ◽  
Jian Yin
Keyword(s):  
Fusion Protein ◽  
Fluorescent Protein ◽  
Expression System ◽  
Baculovirus Expression System ◽  
Size Exclusion ◽  
Coupled Transport ◽  
Gaba Transporter ◽  
Gfp Fusion Protein ◽  
Baculovirus Expression ◽  
Gaba Transporter 1

The γ-aminobutyric acid (GABA) transporter 1 (GAT1) belongs to a family of Na+ and Cl−-coupled transport proteins and possesses 12 putative transmembrane domains. To perform structural analyses of the GAT1 protein, the GAT1/green fluorescent protein (GFP) fusion protein was functionally expressed in insect Sf9 cells by the BAC-TO-BAC® baculovirus expression system. 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 and size-exclusion fast protein liquid chromatography (SE-FPLC). A yield of 200–300 μg of the GAT1/GFP protein could be purified from 400–600 mL of infected Sf9 cells. The purified protein was analyzed by transmission electron microscopy (TEM), which revealed that the GAT1/GFP fusion protein was isolated in its monomeric form.

scholarly journals Expression of functional G protein beta gamma dimers of defined subunit composition using a baculovirus expression system.

1992 ◽  
Vol 267 (19) ◽  
pp. 13123-13126 ◽  
Author(s):  
S.G. Graber ◽  
R.A. Figler ◽  
V.K. Kalman-Maltese ◽  
J.D. Robishaw ◽  
J.C. Garrison
Keyword(s):  
G Protein ◽  
Expression System ◽  
Baculovirus Expression System ◽  
Subunit Composition ◽  
Baculovirus Expression

scholarly journals Production, processing and partial purification of functional G protein βγ subunits in baculovirus-infected insect cells

1992 ◽  
Vol 286 (3) ◽  
pp. 677-680 ◽  
Author(s):  
J D Robishaw ◽  
V K Kalman ◽  
K L Proulx
Keyword(s):  
G Protein ◽  
G Proteins ◽  
Insect Cells ◽  
Expression System ◽  
Baculovirus Expression System ◽  
Partial Purification ◽  
Baculovirus Expression ◽  
Gamma Subunits ◽  
Infected Insect ◽  
Beta 2

As a result of the inability to resolve the heterogeneous mixture of G protein beta gamma subunits present in tissues, it has not been possible to compare different beta gamma subunits of the G proteins in terms of their proposed roles in receptor-effector coupling. This study was undertaken to establish the utility of the baculovirus expression system in producing homogeneous beta gamma subunits of defined composition for the comparative analysis of these subunits in reconstitution systems. In this study we report the expression, and appropriate post-translational processing, of recombinant beta 2, gamma 2 and gamma 3 subunits. In addition, we show that the recombinant beta gamma subunits can be readily purified, and can functionally interact with the alpha subunits of the G proteins.

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