Production of recombinant new canine parvovirus 2a viral protein 2 in SF9 cells using a baculovirus expression system

2020 ◽  
Vol 44 (3) ◽  
pp. 142-145
Author(s):  
Hyeonhae Choi ◽  
Seyeon Park ◽  
Yoon-Hee Lee ◽  
Ju-Yeon Lee ◽  
Jae Young Song ◽  
...  
Keyword(s):  
Viral Protein ◽  
Expression System ◽  
Baculovirus Expression System ◽  
Canine Parvovirus ◽  
Sf9 Cells ◽  
Baculovirus Expression

Expression and characterization of recombinant human α-3/4-fucosyltransferase III from Spodoptera frugiperda (Sf9) and Trichoplusia ni (Tn) cells using the baculovirus expression system

2001 ◽  
Vol 353 (3) ◽  
pp. 719-725 ◽  
Author(s):  
Vanessa A. MORAIS ◽  
Jacinta SERPA ◽  
Angelina S. PALMA ◽  
Teresa COSTA ◽  
Luís MARANGA ◽  
...  
Keyword(s):  
Cell Lines ◽  
Trichoplusia Ni ◽  
Secretory Pathway ◽  
Expression System ◽  
Baculovirus Expression System ◽  
Exchange Chromatography ◽  
Sf9 Cells ◽  
Type I ◽  
Baculovirus Expression ◽  
Insect Cell Lines

The human α-3/4-fucosyltransferase III (Fuc-TIII) participates in the synthesis of Lewis determinants. The enzyme from human sources is scarce and heterogeneous. In this paper we describe the expression of a secreted form of Fuc-TIII (SFT3) in two insect cell lines, Spodopterafrugiperda (Sf9) and Trichoplusiani (Tn), using the baculovirus expression system. The Sf9 cells secreted approx. 0.4unit/l (1mg/l) of the enzyme. The Tn cells secreted approx. 3-fold this amount. A large proportion of active protein was accumulated in the two cell lines (50 and 75% respectively for Sf9 and Tn cells, on the fourth day after infection) indicating a possible limitation not only of the folding machinery, but also a saturation of the secretory pathway. SFT3 was purified by cation-exchange chromatography followed by affinity chromatography. The enzyme from the Tn cell line had a lower global charge, possibly due to post-translational modifications, such as phosphorylation or sulphation. The two glycosylation sites from SFT3 were occupied. SFT3 secreted by Sf9 cells was completely deglycosylated by peptide-N-glycanase F, whereas 50% of SFT3 secreted by Tn cells was resistant to deglycosylation by this enzyme. The apparent kinetic parameters determined with the type I acceptor were kcat = 0.4s-1 and Km = 0.87mM for the SFT3 secreted by Tn cells, and kcat = 0.09s-1 and Km = 0.76mM for the SFT3 secreted by Sf9 cells, indicating that the enzymes had substrate affinities within the same order of magnitude as their mammalian counterpart. Furthermore, SFT3 secreted by either cell type showed a clear preference for type 1 carbohydrate acceptors, similarly to human Fuc-TIII.

Production of VP2 protein of canine parvovirus-2c expressed in baculovirus expression system

2020 ◽  
Vol 44 (2) ◽  
pp. 96-98
Author(s):  
Seyeon Park ◽  
◽  
Ju-Yeon Lee ◽  
Jae Young Song ◽  
Yoon-Hee Lee ◽  
...  
Keyword(s):  
Expression System ◽  
Baculovirus Expression System ◽  
Canine Parvovirus ◽  
Vp2 Protein ◽  
Baculovirus Expression

The C-terminal tail domain of neurofilament protein-H (NF-H) forms the crossbridges and regulates neurofilament bundle formation

2000 ◽  
Vol 113 (21) ◽  
pp. 3861-3869 ◽  
Author(s):  
J. Chen ◽  
T. Nakata ◽  
Z. Zhang ◽  
N. Hirokawa
Keyword(s):  
Network Formation ◽  
Expression System ◽  
Baculovirus Expression System ◽  
Sf9 Cells ◽  
Tail Domain ◽  
Neurofilament Protein ◽  
Baculovirus Expression ◽  
10 Nm Filaments ◽  
Parallel Arrays

In order to study the role of NF-H in a neurofilament network formation in neurons, we coexpressed NF-H with neurofilament protein-L (NF-L) in Sf9 cells using the baculovirus expression system. Electron microscopy observations revealed that parallel arrays of 10 nm filaments with frequent crossbridges between adjacent filaments were formed in the cytoplasm of Sf9 cells infected with the recombinant virus that co-expressed NF-L and NF-H. To explore the function of the C-terminal tail domain of NF-H, various deletion mutants lacking portions of the tail domain were constructed, and each of them was coexpressed with NF-L. The results show that the tail domain of NF-H is a structural component of crossbridges and is involved in parallel bundle formation of neurofilaments, as core filaments of the axon. The last 191 amino acids of the C-terminal tail domain of NF-H play a key role in crossbridge formation.

Functional Expression of Gastric H,K-ATPase in Sf9 Cells using the Baculovirus Expression System

1994 ◽  
pp. 19-26
Author(s):  
Corné H. W. Klaassen ◽  
Tom J. F. Van Uem ◽  
Mariëlle P. De Moel ◽  
Godelieve L. J. De Caluwé ◽  
Herman G. P. Swarts ◽  
...  
Keyword(s):  
Expression System ◽  
Baculovirus Expression System ◽  
Functional Expression ◽  
Sf9 Cells ◽  
Baculovirus Expression

scholarly journals Enhanced production of Canine parvovirus VP2  protein using improved Baculovirus expression  system  and its immunogenicity

2020 ◽  
Author(s):  
Dao Chang ◽  
Yangkun Liu ◽  
Yangyang Chen ◽  
Xiaomin Hu ◽  
Andrey Burov ◽  
...  
Keyword(s):  
Expression System ◽  
Baculovirus Expression System ◽  
Canine Parvovirus ◽  
Full Length ◽  
Recombinant Viruses ◽  
Vp2 Protein ◽  
Enhanced Production ◽  
Baculovirus Expression ◽  
Yield Production ◽  
High Level

Abstract Background : Canine parvovirus (CPV) is now recognized as a serious threat to dog industry worldwide. Vaccination remains the principal tool to control CPV infection. However, due to low yield, production of VP2 protein of CPV in baculovirus expression system remains challenging. The aim of this study was to increase the VP2 protein production by using a improved baculovirus expression system (Multibac) and evaluate the immunogenicity of the purified VP2 protein in mice. Results: The results showed that CPV VP2 protein was successfully expressed in the improved baculovirus expression system efficiently. A high level of expression of the full length VP2 protein was achieved using our modified system. The recombinant virus carrying two copies of VP2 showed the highest expression level, with a productivity of 186 mg/L, which is about 1.4-1.6 fold that of the recombinant viruses carrying only one copy. The purified protein could react with Mouse anti-His tag monoclonal antibody and Rabbit anti-VP2 polyclonal antibody with good reactogenicity. The mice were then immunized with purified full length VP2 protein to evaluate its immunogenicity. After vaccination, VP2 protein could induce the mice produce high level of hemagglutination inhibition antibodies. Conclusions: Full length CPV VP2 protein was successfully expressed at high level and purified efficiently. And it stimulated mice to produce high level of antibody. The full length VP2 expressed in this study could be used as a putative economic and efficient subunit vaccine against CPV infection.

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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