Functional Expression of theAequorea victoriaGreen Fluorescent Protein in Insect Cells Using the Baculovirus Expression System

1996 ◽  
Vol 219 (1) ◽  
pp. 14-20 ◽  
Author(s):  
Helmut Reiländer ◽  
Winfried Haase ◽  
Gabi Maul
Keyword(s):  
Insect Cells ◽  
Fluorescent Protein ◽  
Expression System ◽  
Baculovirus Expression System ◽  
Functional Expression ◽  
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.

scholarly journals Partial characterization of natural and recombinant human soluble CD23

1992 ◽  
Vol 286 (3) ◽  
pp. 819-824 ◽  
Author(s):  
K Rose ◽  
G Turcatti ◽  
P Graber ◽  
S Pochon ◽  
P O Regamey ◽  
...  
Keyword(s):  
Insect Cells ◽  
Peptide Mapping ◽  
Sequence Similarity ◽  
Expression System ◽  
Protein A ◽  
Baculovirus Expression System ◽  
Partial Characterization ◽  
Disulphide Bonds ◽  
Baculovirus Expression

The purification to homogeneity of an active soluble 25 kDa fragment of CD23, produced in insect cells using the baculovirus expression system, is described. Peptide mapping and analysis by Edman degradation and mass spectrometry permitted partial characterization of the protein. A total of 165 out of 172 residues, including N-terminal and C-terminal regions, were mapped. The positions of the two disulphide bonds in the IgE-binding region were also determined: residue 110 is joined to residue 124, and residue 42 to residue 133. Natural CD23 25 kDa fragment was also analysed and found to possess the same disulphide bond arrangement. These results extend the previously noted sequence similarity with lectins to elements of secondary structure.

Strong buffering capacity of insect cells. Implications for the baculovirus expression system

1995 ◽  
Vol 17 (1) ◽  
pp. 21-26 ◽  
Author(s):  
Miguel Medina ◽  
Abelardo L�pez-Rivas ◽  
Douwe Zuidema ◽  
Graham J. Belsham ◽  
Esteban Domingo ◽  
...  
Keyword(s):  
Insect Cells ◽  
Expression System ◽  
Baculovirus Expression System ◽  
Buffering Capacity ◽  
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 Synthesis of tetanus toxin fragment C in insect cells by use of a baculovirus expression system.

1991 ◽  
Vol 59 (5) ◽  
pp. 1627-1632 ◽  
Author(s):  
I G Charles ◽  
B C Rodgers ◽  
A J Makoff ◽  
S N Chatfield ◽  
D E Slater ◽  
...  
Keyword(s):  
Tetanus Toxin ◽  
Insect Cells ◽  
Expression System ◽  
Baculovirus Expression System ◽  
Baculovirus Expression

scholarly journals Enhanced Production of Recombinant Protein by Fusion Expression with Ssp DnaB Mini-Intein in the Baculovirus Expression System

Viruses ◽  
2018 ◽  
Vol 10 (10) ◽  
pp. 523 ◽  
Author(s):  
Won Gwak ◽  
Jae Choi ◽  
Beom Han ◽  
Sung Bae ◽  
Soo Woo
Keyword(s):  
Fluorescent Protein ◽  
Expression System ◽  
Classical Swine Fever ◽  
Baculovirus Expression System ◽  
Additional Treatment ◽  
Fusion Partner ◽  
Target Proteins ◽  
Enhanced Production ◽  
Baculovirus Expression ◽  
Green Fluorescent

The baculovirus expression system (BES) is considered to be a very powerful tool for the expression of numerous difficult to express vertebrate proteins. Ssp DnaB mini-intein is a useful fusion partner for the production of recombinant proteins because it can be self-cleaved by controlling the pH and temperature, without additional treatment. To evaluate the utility of Ssp DnaB mini-intein in the BES, recombinant viruses were generated to express the enhanced green fluorescent protein, the VP2 protein of porcine parvovirus, and the E2 protein of classical swine fever virus fused to a mini-intein. As expected, intracellular self-cleavage of the mini-intein occurred during virus infection, but the cleavage initiation time varied depending on the target protein. Significantly enhanced protein production was observed for all of the target proteins that were fused to the mini-intein. This increase was enough to overcome the decrease in the fusion protein due to intracellular self-cleavage. The mini-intein in all of the recombinant fusion proteins was successfully cleaved by controlling the pH and temperature. These results suggest that the Ssp DnaB mini-intein is a useful fusion partner in the BES for easy purification and enhanced production of target proteins.

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