Recombinant baculovirus-based multiple protein expression platform for Drosophila S2 cell culture

2008 ◽  
Vol 133 (1) ◽  
pp. 116-122 ◽  
Author(s):  
Kyoung Ro Kim ◽  
Yeon Kyu Kim ◽  
Hyung Joon Cha
Keyword(s):  
Cell Culture ◽  
Protein Expression ◽  
Recombinant Baculovirus ◽  
Expression Platform ◽  
Multiple Protein

MO710GALECTIN-3, IS IT A NEW PLAYER IN PERITONEAL INFLAMMATION AMONG PATIENTS UNDERGOING PERITONEAL DIALYSIS?

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
Yael Einbinder ◽  
Keren Cohen-Hagai ◽  
Sydney Benchetrit ◽  
Tali Zitman-Gal
Keyword(s):  
Cell Culture ◽  
Peritoneal Dialysis ◽  
Endothelial Cell ◽  
Protein Expression ◽  
Inflammatory Process ◽  
Peritoneal Membrane ◽  
Peritoneal Equilibration Test ◽  
Endothelial Cell Culture ◽  
Dialysis Vintage ◽  
Mrna And Protein Expression

Abstract Background and Aims Peritoneal dialysis (PD) is a common used method for renal replacement therapy. Prolonged PD treatment causes structural and functional changes in the peritoneal membrane which are attributed to local inflammatory process in the peritoneal cavity. Galectin-3 (Gal-3) is a galactoside-binding lectin with pro-inflammatory and pro-fibrotic effects. The aim of this study was to assess correlation between Gal-3 serum and dialysate effluent levels with peritoneal membrane transport characteristics. Method Gal-3 levels in serum and dialysate effluent were measured simultaneously in prevalent PD patients in morning visit or during peritoneal equilibration test (PET). Gal-3 levels were correlated with clinical and laboratory parameters. Interlukin (IL) -6 levels were measured in dialysate effluent. Gal-3 mRNA and protein expression were evaluated after exposure of primary endothelial cell culture to several dialysate solutions. Results 37 PD patients were included in the study; mean age was 65.7±13.1 years, mean dialysis vintage was 17.5±13 months. Gal-3 levels in dialysate effluent correlated with peritoneal equilibration test (PET) results (0.663, p=0.005) and effluent IL-6 levels (0.674, p=0.002) but not with serum Gal-3 levels or dialysis vintage. Patients with high PET results had higher effluent Gal-3 levels as compared average low PET results. In multivariate regression analysis effluent IL-6 level was the most dominant predictor of effluent Gal-3 levels. Gal-3 mRNA and protein expression in primary endothelial cell culture were not affected by stimulation with dialysate solutions. Conclusion Our study demonstrated presence of Gal-3 within the dialysate effluent in PD patients. Gal-3 levels correlated with peritoneal membrane transport characteristics and effluent IL-6 levels suggesting a role in the inflammatory process within the peritoneal cavity.

Cell Culture Block Array for Immunocytochemical Study of Protein Expression in Cultured Cells

2005 ◽  
Vol 13 (1) ◽  
pp. 85-90 ◽  
Author(s):  
Rongshan Li ◽  
Jing Ni ◽  
Patricia A. Bourne ◽  
Shuyuan Yeh ◽  
Jorge Yao ◽  
...  
Keyword(s):  
Cell Culture ◽  
Protein Expression ◽  
Cultured Cells ◽  
Immunocytochemical Study

scholarly journals Inhibition of human cytomegalovirus immediate-early gene expression by an antisense oligonucleotide complementary to immediate-early RNA.

1996 ◽  
Vol 40 (9) ◽  
pp. 2004-2011 ◽  
Author(s):  
K P Anderson ◽  
M C Fox ◽  
V Brown-Driver ◽  
M J Martin ◽  
R F Azad
Keyword(s):  
Gene Expression ◽  
Cell Culture ◽  
Antiviral Activity ◽  
Protein Expression ◽  
Human Cytomegalovirus ◽  
Host Cells ◽  
Early Gene ◽  
Immediate Early ◽  
Mrna Levels ◽  
Virus Adsorption

ISIS 2922 is a phosphorothioate oligonucleotide that is complementary to human cytomegalovirus (CMV) immediate-early (IE) RNA and that exhibits potent and specific antiviral activity against CMV in cell culture assays. Specific assay systems were developed to separately characterize the antisense and nonantisense components of the antiviral activity mediated by ISIS 2922. In U373 cells transformed with cDNA encoding the CMV IE 55-kDa (IE55) protein, expression was inhibited at nanomolar concentrations comparable to effective concentrations in antiviral assays. The specificity of inhibition was demonstrated by using control oligonucleotides incorporating progressive base changes to destabilize oligonucleotide-RNA base pairing and by showing a lack of inhibition of the CMV IE72 product expressed from the same promoter. Inhibition of IE55 protein expression correlated with a reduction in mRNA levels consistent with an RNase H-mediated termination event. Studies with virus-infected cells demonstrated that antisense and nonantisense mechanisms contribute to the antiviral activity of ISIS 2922. Base complementarity to target RNA was important for optimal activity in antiviral assays, but base changes affecting parameters other than hybridization affinity also influenced antiviral activity. Sequence-independent inhibition of virus adsorption to host cells by phosphorothioate oligonucleotides was also observed at high concentrations. Therefore, at least three different mechanisms may contribute to the antiviral activity of ISIS 2922 in cell culture: antisense-mediated inhibition of target gene expression; nonantisense, sequence-dependent inhibition of virus replication; and sequence-independent inhibition of virus adsorption to host cells.

scholarly journals Generation of Infectious Hepatitis C Virus in Immortalized Human Hepatocytes

2006 ◽  
Vol 80 (9) ◽  
pp. 4633-4639 ◽  
Author(s):  
Tatsuo Kanda ◽  
Arnab Basu ◽  
Robert Steele ◽  
Takaji Wakita ◽  
Jan S. Ryerse ◽  
...  
Keyword(s):  
Cell Culture ◽  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Protein Expression ◽  
Culture Media ◽  
Human Hepatocytes ◽  
Full Length ◽  
Dependent Manner ◽  
Virus Growth ◽  
Hcv Genotype

ABSTRACT Progress in understanding hepatitis C virus (HCV) biology has remained a challenge due to the lack of an efficient cell culture system for virus growth. In this study, we examined HCV core protein-mediated immortalized human hepatocytes (IHH) for growth of HCV. In vitro-transcribed full-length RNA from HCV genotype 1a (clone H77) was introduced into IHH by electroporation. Reverse transcription-PCR of cellular RNA isolated from HCV genome-transfected IHH suggested that viral RNA replication occurred. IHH transfected with the full-length HCV genome also displayed viral protein expression by indirect immunofluorescence. In contrast, cells transfected with polymerase-defective HCV (H77/GND) RNA as a negative control did not exhibit expression of the viral genome. Immunogold labeling demonstrated localization of E1 protein in the rough endoplasmic reticulum of RNA-transfected IHH. Virus-like particles of ∼50 nm were observed in the cytoplasm. After being inoculated with culture media of cells transfected with the full-length HCV genome, naïve IHH displayed NS5a protein expression in a dilution-dependent manner, but expression of NS5a was inhibited by prior incubation of culture medium with HCV-infected patient sera. NS5a-positive immunofluorescence of cell culture media of IHH transfected with full-length H77 RNA yielded ∼4.5 × 104 to 1 × 105 focus-forming units/ml. A similar level of virus growth was observed upon transfection of RNA from HCV genotype 2a (JFH1) into IHH. Taken together, our results suggest that IHH support HCV genome replication and virus assembly.

Orthogonal, light-inducible protein expression platform in yeast Sacchararomyces cerevisiae

2018 ◽  
Vol 44 ◽  
pp. S19
Author(s):  
K. Messerschmidt ◽  
F. Machens ◽  
L. Hochrein ◽  
G. Naseri
Keyword(s):  
Protein Expression ◽  
Expression Platform ◽  
Inducible Protein

scholarly journals Application of the LEXSY Leishmania tarentolae system as a recombinant protein expression platform: A review

2019 ◽  
Vol 87 ◽  
pp. 164-173
Author(s):  
Tatiana Aparecida de Oliveira ◽  
Walmir da Silva ◽  
Nancy da Rocha Torres ◽  
João Victor Badaró de Moraes ◽  
Renato Lima Senra ◽  
...  
Keyword(s):  
Recombinant Protein ◽  
Protein Expression ◽  
Recombinant Protein Expression ◽  
Leishmania Tarentolae ◽  
Expression Platform

scholarly journals NSm and 78-Kilodalton Proteins of Rift Valley Fever Virus Are Nonessential for Viral Replication in Cell Culture

2006 ◽  
Vol 80 (16) ◽  
pp. 8274-8278 ◽  
Author(s):  
Sungyong Won ◽  
Tetsuro Ikegami ◽  
C. J. Peters ◽  
Shinji Makino
Keyword(s):  
Cell Culture ◽  
Protein Expression ◽  
Growth Kinetics ◽  
Rift Valley ◽  
Rift Valley Fever ◽  
Rift Valley Fever Virus ◽  
Parent Virus ◽  
M Gene ◽  
Valley Fever ◽  
Similar Growth

ABSTRACT Rift Valley fever viruses carrying mutations of the M gene preglycoprotein region, one lacking NSm protein expression, one lacking 78-kDa protein expression, and one lacking expression of both proteins, were compared in cell culture. All of the mutants and their parent virus produced plaques with similar sizes and morphologies in Vero E6 cells and had similar growth kinetics in Vero, C6/36, and MRC5 cells, demonstrating that the NSm and 78-kDa proteins were not needed for the virus to replicate efficiently in cell culture. A competition-propagation assay revealed that the parental virus was slightly more fit than the mutant virus lacking expression of both proteins.

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