The Lentiviral System Construction for Highly Expressed Porcine Stearoyl-CoA Desaturase-1 and Functional Characterization in Stably Transduced Porcine Swine Kidney Cells

Lipids ◽  
2018 ◽  
Vol 53 (10) ◽  
pp. 933-945
Author(s):  
Jinhee Hwang ◽  
Neetu Singh ◽  
Charles Long ◽  
Stephen B. Smith
Keyword(s):  
Functional Characterization ◽  
Kidney Cells ◽  
System Construction ◽  
Stearoyl Coa Desaturase ◽  
Swine Kidney

scholarly journals Hepatitis E genotype 3 virus isolate from wild boar is capable of replication in non-human primate and swine kidney cells and mouse neuroblastoma cells

2020 ◽  
Vol 16 (1) ◽  
Author(s):  
Juozas Grigas ◽  
Evelina Simkute ◽  
Martynas Simanavicius ◽  
Arnoldas Pautienius ◽  
Zaneta Streimikyte-Mockeliune ◽  
...  
Keyword(s):  
Wild Boar ◽  
Virus Isolate ◽  
Neuroblastoma Cells ◽  
Hepatitis E ◽  
Kidney Cells ◽  
Genotype 3 ◽  
Mouse Neuroblastoma ◽  
Swine Kidney ◽  
Human Primate

scholarly journals Inactivation of the RNase Activity of Glycoprotein Erns of Classical Swine Fever Virus Results in a Cytopathogenic Virus

1998 ◽  
Vol 72 (1) ◽  
pp. 151-157 ◽  
Author(s):  
M. M. Hulst ◽  
F. E. Panoto ◽  
A. Hoekman ◽  
H. G. P. van Gennip ◽  
R. J. M. Moormann
Keyword(s):  
Virus Infection ◽  
Classical Swine Fever Virus ◽  
Polyclonal Antibodies ◽  
Classical Swine Fever ◽  
Fever Virus ◽  
Rnase Activity ◽  
Kidney Cells ◽  
Parent Virus ◽  
Induced Apoptosis ◽  
Swine Kidney

ABSTRACT Envelope glycoprotein Erns of classical swine fever virus (CSFV) has been shown to contain RNase activity and is involved in virus infection. Two short regions of amino acids in the sequence of Erns are responsible for RNase activity. In both regions, histidine residues appear to be essential for catalysis. They were replaced by lysine residues to inactivate the RNase activity. The mutated sequence of Erns was inserted into the p10 locus of a baculovirus vector and expressed in insect cells. Compared to intact Erns, the mutated proteins had lost their RNase activity. The mutated proteins reacted with Erns-specific neutralizing monoclonal and polyclonal antibodies and were still able to inhibit infection of swine kidney cells (SK6) with CSFV, but at a concentration higher than that measured for intact Erns. This result indicated that the conformation of the mutated proteins was not severely affected by the inactivation. To study the effect of these mutations on virus infection and replication, a CSFV mutant with an inactivated Erns (FLc13) was generated with an infectious DNA copy of CSFV strain C. The mutant virus showed the same growth kinetics as the parent virus in cell culture. However, in contrast to the parent virus, the RNase-negative virus induced a cytopathic effect in swine kidney cells. This effect could be neutralized by rescue of the inactivated Erns gene and by neutralizing polyclonal antibodies directed against Erns, indicating that this effect was an inherent property of the RNase-negative virus. Analyses of cellular DNA of swine kidney cells showed that the RNase-negative CSFV induced apoptosis. We conclude that the RNase activity of envelope protein Erns plays an important role in the replication of pestiviruses and speculate that this RNase activity might be responsible for the persistence of these viruses in their natural host.

scholarly journals Hepatitis E Virus Genotype 1 Infection of Swine Kidney Cells In Vitro Is Inhibited at Multiple Levels

2013 ◽  
Vol 88 (2) ◽  
pp. 868-877 ◽  
Author(s):  
H. T. Nguyen ◽  
P. Shukla ◽  
U. Torian ◽  
K. Faulk ◽  
S. U. Emerson
Keyword(s):  
Hepatitis E Virus ◽  
Hepatitis E ◽  
Kidney Cells ◽  
Genotype 1 ◽  
Virus Genotype ◽  
Multiple Levels ◽  
Swine Kidney

Temporary disturbance of actin stress fibers in swine kidney cells during pseudorabies virus infection

2002 ◽  
Vol 86 (1-2) ◽  
pp. 89-94 ◽  
Author(s):  
G Van Minnebruggen ◽  
G.R Van de Walle ◽  
H.W Favoreel ◽  
H.J Nauwynck ◽  
M.B Pensaert
Keyword(s):  
Virus Infection ◽  
Pseudorabies Virus ◽  
Stress Fibers ◽  
Kidney Cells ◽  
Temporary Disturbance ◽  
Swine Kidney ◽  
Actin Stress Fibers

scholarly journals Passage of Classical Swine Fever Virus in Cultured Swine Kidney Cells Selects Virus Variants That Bind to Heparan Sulfate due to a Single Amino Acid Change in Envelope Protein Erns

2000 ◽  
Vol 74 (20) ◽  
pp. 9553-9561 ◽  
Author(s):  
M. M. Hulst ◽  
H. G. P. van Gennip ◽  
R. J. M. Moormann
Keyword(s):  
Amino Acid ◽  
Cell Surface ◽  
Heparan Sulfate ◽  
Classical Swine Fever Virus ◽  
Classical Swine Fever ◽  
Kidney Cells ◽  
Virus Particles ◽  
Heparinase I ◽  
Initial Binding ◽  
Swine Kidney

ABSTRACT Infection of cells with Classical swine fever virus (CSFV) is mediated by the interaction of envelope glycoprotein Ernsand E2 with the cell surface. In this report we studied the role of the cell surface glycoaminoglycans (GAGs), chondroitin sulfates A, B, and C (CS-A, -B, and -C), and heparan sulfate (HS) in the initial binding of CSFV strain Brescia to cells. Removal of HS from the surface of swine kidney cells (SK6) by heparinase I treatment almost completely abolished infection of these cells with virus that was extensively passaged in swine kidney cells before it was cloned (clone C1.1.1). Infection with C1.1.1 was inhibited completely by heparin (a GAG chemically related to HS but sulfated to a higher extent) and by dextran sulfate (an artificial highly sulfated polysaccharide), whereas HS and CS-A, -B, and -C were unable to inhibit infection. Bound C1.1.1 virus particles were released from the cell surface by treatment with heparin. Furthermore, C1.1.1 virus particles and CSFV Ernspurified from insect cells bound to immobilized heparin, whereas purified CSFV E2 did not. These results indicate that initial binding of this virus clone is accomplished by the interaction of Erns with cell surface HS. In contrast, infection of SK6 cells with virus clones isolated from the blood of an infected pig and minimally passaged in SK6 cells was not affected by heparinase I treatment of cells and the addition of heparin to the medium. However, after one additional round of amplification in SK6 cells, infection with these virus clones was affected by heparinase I treatment and heparin. Sequence analysis of the Erns genes of these virus clones before and after amplification in SK6 cells showed that passage in SK6 cells resulted in a change of an Ser residue to an Arg residue in the C terminus of Erns (amino acid 476 in the polyprotein of CSFV). Replacement of the Erns gene of an infectious DNA copy of C1.1.1 with the Erns genes of these virus variants proved that acquisition of this Arg was sufficient to alter an HS-independent virus to a virus that uses HS as an Erns receptor.

Structured Matrices Associated With Adenovirus-Infected Swine Kidney Cells

1968 ◽  
Vol 26 ◽  
pp. 28-29
Author(s):  
A. E. Ritchie ◽  
J. O. Norman
Keyword(s):  
Cell Culture ◽  
Distilled Water ◽  
Kidney Cells ◽  
Special Relationship ◽  
Structured Matrices ◽  
Cell Monolayers ◽  
Carbon Coated ◽  
Infected Cells ◽  
Viral Maturation ◽  
Swine Kidney

Fibrillar matrices (viroplasms) have been described for a number of viruses, mainly from studies of thin-sectioned infected cells. We have recently noted some structured matrices (observed in negatively-stained preparations) that were associated with swine kidney (SK) cells infected with an Adenovirus, infectious canine hepatitis. These cellular fragments may bear some special relationship to the process of viral maturation.Virus was propagated in primary swine kidney cell monolayers. Cell culture harvests were pooled and clarified by centrifugation at 1000 X g for 20 min at 4 C. The pelleted materials were dispersed in 5 to 10 times their volume of distilled water and sampled directly for phosphotungstate (PTA) negative staining: one drop of the suspension was diluted with ca. 15 drops of 1% PTA which contained a trace of bovine serum albumin (Fr. V); the sample was then applied to carbon-coated collodion-filmed grids with a Vaponefrin all-glass nebulizer (Brenner & Horne. Biochim. Biophys. Acta 34: 103 (1959). The process of nebulizing appeared to rupture those cells that reached the specimen grid permitting examination of cellular fragments in great variety.

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