Cloning and expression of a putative glycoprotein gene of channel catfish virus using baculovirus expression system

The channel catfish virus (CCV, Ictalurid herpesvirus 1) has caused sustained economic losses in the fish industry because of its strong infectivity and pathogenicity. Thus, it is necessary to determine the function of viral proteins in the CCV infection process. The present study aimed to characterize CCV glycoprotein ORF59 and explore its impact on virus infection in host cells. Firstly, its exclusive presence in the membrane fraction of the cell lysate and subcellular localization verified that CCV ORF59 is a viral membrane protein expressed at late-stage infection. A protein blocking assay using purified His6 tagged ORF59, expressed in sf9 insect cells using a baculovirus expression system, indicated a dose-dependent inhibitory effect of recombinant ORF59 protein on virus invasion. Knockdown of the ORF59 using a short hairpin (shRNA) showed that ORF59 silencing decreased the production of infectious virus particles in channel catfish ovary cells. The results of this study suggest that recombinant ORF59 protein might inhibit CCV entry into the host cells. These findings will promote future studies of the key functions of glycoprotein ORF59 during CCV infection.

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Cloning and expression of a cellulase gene in the silkworm, Bombyx mori by improved Bac-to-Bac/BmNPV baculovirus expression system

Molecular Biology Reports ◽

10.1007/s11033-010-0025-2 ◽

2010 ◽

Vol 37 (8) ◽

pp. 3721-3728 ◽

Cited By ~ 11

Author(s):

Xing-hua Li ◽

Dan Wang ◽

Fang Zhou ◽

Hua-jun Yang ◽

Roy Bhaskar ◽

...

Keyword(s):

Bombyx Mori ◽

Expression System ◽

Baculovirus Expression System ◽

Cloning And Expression ◽

Cellulase Gene ◽

Baculovirus Expression ◽

Silkworm Bombyx Mori

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

Journal of Biological Chemistry ◽

10.1016/s0021-9258(18)42178-3 ◽

1992 ◽

Vol 267 (19) ◽

pp. 13123-13126 ◽

Cited By ~ 4

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

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

Biochemical Journal ◽

10.1042/bj2860677 ◽

1992 ◽

Vol 286 (3) ◽

pp. 677-680 ◽

Cited By ~ 21

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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