Journal: Journal of Molecular Biology

Article Title: Display of epitopes on the surface of tobacco mosaic virus: impact of charge and isoelectric point of the epitope on virus-host interactions 1

doi: 10.1006/jmbi.1999.2860

Figure Lengend Snippet: Accumulation of (a) basic and (b) acidic PR2 mRNA in N. tabacum Xanthi-nn and Xanthi-NN plants following inoculation with TMV-U1 or TMV-RB19 at 4 d.p.i. RNA dot blots were performed using similar amounts of total RNA and then hybridized with sequences to detect (a) basic or (b) acidic PR2 gene expression. The signal intensity of the isotope label was measured using phosphoimager followed by Multi-Analyst computer program analyses (Bio-Rad, Hercules, CA), and the relative values are displayed as graphs.

Article Snippet: The signal intensity of the isotope label was measured using phosphoimager followed by Multi-Analyst computer program analyses (Bio-Rad, Hercules, CA), and the relative values are displayed as graphs.

Techniques: Gene Expression

Journal: Journal of Virology

Article Title: The NS5A/NS5 Proteins of Viruses from Three Genera of the Family Flaviviridae Are Phosphorylated by Associated Serine/Threonine Kinases

doi: 10.1128/jvi.72.7.6199-6206.1998

Figure Lengend Snippet: FIG. 6. Comparison of the effects of divalent cation concentration on the in vitro phosphorylation of GST-BVDV NS5A, GST-YF NS5, and GST-HCV NS5A. pTM3/GST-BVDV 5A, pBRTM/GST-YF 5, or pTM3/GST-HCV 5A was expressed in BHK-21 cells, and the respective fusion proteins were purified and assayed for associated kinase activity as previously described (36). Clarified lysates were pooled and divided into equal aliquots prior to isolation on glutathione-agarose to ensure that all in vitro kinase reactions contained equal amounts of protein. Standard kinase wash buffers and kinase reaction buffers were used in these experiments with the following exceptions: (i) 5 mM MnCl2 was replaced with the indicated concentrations of MnCl2 in reaction mixtures for which activities are shown in panels on the left, and (ii) the indicated concentrations of CaCl2 were included, along with 5 mM MnCl2, in reaction mixtures for which activities are shown in the panels on the right. The level of phosphorylation in each reaction was determined by phosphorimager quantitation (Bio-Rad) of SDS-8% polyacrylamide gels.

Article Snippet: The level of phosphorylation in each reaction was determined by phosphorimager quantitation (Bio-Rad) of SDS-8% polyacrylamide gels. on S eptem ber 13, 2020 by guest http://jvi.asm .org/ D ow nloaded from conservation of these structural elements may reflect their involvement in some function common to the NS5 and NS5A proteins of the Flaviviridae which has been conserved throughout their evolution from a common ancestor.

Techniques: Comparison, Concentration Assay, In Vitro, Phospho-proteomics, Activity Assay, Isolation, Quantitation Assay

Journal: Journal of Virology

Article Title: The NS5A/NS5 Proteins of Viruses from Three Genera of the Family Flaviviridae Are Phosphorylated by Associated Serine/Threonine Kinases

doi: 10.1128/jvi.72.7.6199-6206.1998

Figure Lengend Snippet: FIG. 7. Comparison of the effects of protein kinase inhibitors on GST- BVDV NS5A, GST-YF NS5, and GST-HCV NS5A phosphorylation in vitro. Assays were performed as previously described (36), except that the kinase reaction buffers contained various concentrations of protein kinase inhibitors, as indicated in micromolar units. Provided below each lane are the percentages of NS5A or NS5 phosphorylation, as determined by phosphorimager quantitation, relative to the appropriate solvent controls (dimethyl sulfoxide [DMSO] for bisindolylmaleimide I-HCl [B], H-89, olomoucine [O], and staurosporine [S] and ethanol [EtOH] for DRB). An additional dimethyl sulfoxide control is shown for GST-HCV NS5A phosphorylation assays containing staurosporine because these samples were analyzed on a separate SDS-8% polyacrylamide gel.

Article Snippet: The level of phosphorylation in each reaction was determined by phosphorimager quantitation (Bio-Rad) of SDS-8% polyacrylamide gels. on S eptem ber 13, 2020 by guest http://jvi.asm .org/ D ow nloaded from conservation of these structural elements may reflect their involvement in some function common to the NS5 and NS5A proteins of the Flaviviridae which has been conserved throughout their evolution from a common ancestor.

Techniques: Comparison, Phospho-proteomics, In Vitro, Quantitation Assay, Solvent, Control