bvdv nadl strain  (ATCC)


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    Name:
    Bovine viral diarrhea virus 1
    Description:
    Applications REFERENCE STRAIN Western Hemisphere Committee on Non primate Animal Virus Characterization WHO
    Catalog Number:
    VR-534
    Price:
    None
    Applications:
    REFERENCE STRAIN: Western Hemisphere Committee on Non-primate Animal Virus Characterization (WHO).
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    Structured Review

    ATCC bvdv nadl strain
    Antibody neutralization against <t>BVDV</t> viral infection. (A) Normal MDBK cells grown in cell culture medium. (B) MDBK cells incubated with BVDV <t>NADL</t> virus pretreated with the serum sample (1:320 dilution) pooled from mice immunized with the FanC-STa-E2 fusion.
    Applications REFERENCE STRAIN Western Hemisphere Committee on Non primate Animal Virus Characterization WHO
    https://www.bioz.com/result/bvdv nadl strain/product/ATCC
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    bvdv nadl strain - by Bioz Stars, 2021-09
    94/100 stars

    Images

    1) Product Images from "A Multiepitope Fusion Antigen Elicits Neutralizing Antibodies against Enterotoxigenic Escherichia coli and Homologous Bovine Viral Diarrhea Virus In Vitro"

    Article Title: A Multiepitope Fusion Antigen Elicits Neutralizing Antibodies against Enterotoxigenic Escherichia coli and Homologous Bovine Viral Diarrhea Virus In Vitro

    Journal: Clinical and Vaccine Immunology : CVI

    doi: 10.1128/CVI.00249-13

    Antibody neutralization against BVDV viral infection. (A) Normal MDBK cells grown in cell culture medium. (B) MDBK cells incubated with BVDV NADL virus pretreated with the serum sample (1:320 dilution) pooled from mice immunized with the FanC-STa-E2 fusion.
    Figure Legend Snippet: Antibody neutralization against BVDV viral infection. (A) Normal MDBK cells grown in cell culture medium. (B) MDBK cells incubated with BVDV NADL virus pretreated with the serum sample (1:320 dilution) pooled from mice immunized with the FanC-STa-E2 fusion.

    Techniques Used: Neutralization, Infection, Cell Culture, Incubation, Mouse Assay

    Related Articles

    other:

    Article Title: Bovine Viral Diarrhea Virus (BVDV): A Preliminary Study on Antiviral Properties of Some Aromatic and Medicinal Plants
    Article Snippet: Viruses representative of ssRNA+, of ssRNA-, and of DNA virus were, respectively, (i) yellow fever virus (YFV) (strain 17-D vaccine (Stamaril Pasteur J07B01)), bovine viral diarrhea virus (BVDV) (strain NADL (ATCC VR-534)), west Nile virus (WNV) (clinical isolate), dengue virus (DENV-2) (clinical isolate), coxsackie type B5 (CV-B5), strain Faulkner (ATCC VR-185), and poliovirus type-1 (Sb-1), Sabin strain Chat (ATCC VR-1562); (ii) human respiratory syncytial virus (hRSV) (strain A2 (ATCC VR-1540)) and vesicular stomatitis virus (VSV) (lab strain Indiana (ATCC VR 1540)); and (iii) vaccinia virus (VV) (vaccine strain Elstree-Lister (ATCC VR-1549)) and human herpes 1 (HSV-1) (strain KOS (ATCC VR- 1493)).

    Article Title: PD-1-Mediated PI3K/Akt/mTOR, Caspase 9/Caspase 3 and ERK Pathways Are Involved in Regulating the Apoptosis and Proliferation of CD4+ and CD8+ T Cells During BVDV Infection in vitro
    Article Snippet: The CP BVDV-1a (strain NADL, No. VR-534) was from the American Type Culture Collection (ATCC).

    Article Title: Potent and Selective Activity against Human Immunodeficiency Virus 1 (HIV-1) of Thymelaea hirsuta Extracts
    Article Snippet: Mutants carrying NRTI mutations, such as the AZTR strain (67N, 70R, 215F, 219Q) and the MDR strain (74V, 41L, 106A, 215Y) were also tested; (ii) Flaviviridae: bovine viral diarrhea virus (BVDV) (strain NADL (ATCC VR-534)); (iii) Picornaviridae: human enterovirus C (poliovirus type-1 (Sb-1), Sabin strain Chat (ATCC VR-1562)).

    Article Title: Antiviral effects of nisin, lysozyme, lactoferrin and their mixtures against bovine viral diarrhoea virus
    Article Snippet: The cytopathic strain of bovine viral diarrhoea virus 1 (NADL strain, ATCC VR-534) was propagated and titrated in MDBK cells and stored at − 80 °C until use.

    Infection:

    Article Title: In Vitro Antimicrobial Activity of Essential Oil Extracted from Leaves of Leoheo domatiophorus Chaowasku, D.T. Ngo and H.T. Le in Vietnam
    Article Snippet: .. The following viruses were purchased from the American Type Culture Collection (ATCC), with the exception of Human Immunodeficiency Virus type-1 (HIV-1) and yellow fever virus (YFV):IIIB laboratory strain of HIV-1, was obtained from the supernatant of the persistently infected H9/IIIB cells (NIH 1983); yellow fever virus (YFV) (strain 17-D vaccine (Stamaril Pasteur J07B01)); bovine viral diarrhoea virus (BVDV) (strain NADL (ATCC VR-534)); coxsackie type B4 (CV-B4) (strain J.V.B. (ATCC VR-184)); human enterovirus C (poliovirus type-1 (Sb-1) (Sabin strain Chat (ATCC VR-1562)); vesicular stomatitis virus (VSV) (lab strain Indiana (ATCC VR 1540)); human respiratory syncytial virus (hRSV) (strain A2 (ATCC VR-1540)); reovirus type-1 (Reo-1) (simian virus 12, strain 3651 (ATCC VR- 214)), vaccinia virus (VV) (vaccine strain Elstree-Lister (ATCC VR-1549)); human herpes 1 (HSV-1) (strain KOS (ATCC VR-1493)). ..

    Article Title: Design, Synthesis, Antiviral Evaluation, and SAR Studies of New 1-(Phenylsulfonyl)-1H-Pyrazol−4-yl-Methylaniline Derivatives
    Article Snippet: .. Baby Hamster Kidney (BHK-21) [ATCC CCL10 (C-13) Mesocricetus auratus ] cells were used for the replication of YFV [strain 17-D vaccine (Stamaril Pasteur J07B01)], DENV-2 [clinical isolate], WNV [clinical isolate], and reovirus type-1 (Reo-1) [simian virus 12, strain 3651 (ATCC VR- 214)]; Madin Darby Bovine Kidney (MDBK) [ATCC CCL22 (NBL-1) Bos Taurus ] cells for bovine viral diarrhea virus (BVDV) [strain NADL (ATCC VR-534)]; Monkey kidney (Vero 76) [ATCCCRL 1587 Cercopithecus Aethiops ] cells for human respiratory syncytial virus (RSV) [strain A2 (ATCC VR-1540)], human enterovirus B [coxsackie type B5 (CV-B5) [strain Faulkner (ATCC VR-185)], human enterovirus C [poliovirus type-1 (Sb-1), Sabin strain Chat (ATCC VR-1562)], vesicular stomatitis virus (VSV) [lab strain Indiana (ATCC VR 158)], vaccinia virus (VV) [strain Elstree (Lister Vaccine) (ATCC VR-1549)], and human herpesvirus 1 (HSV-1) [strain KOS (ATCC VR- 1493)]; CD4+ human T-cells containing an integrated HTLV-1 genome (MT-4) for the IIIB laboratory strain of HIV-1, obtained from the supernatant of the persistently infected H9/IIIB cells (NIH 1983). ..

    Article Title: Biological Activities of Essential Oils from Leaves of Paramignya trimera (Oliv.) Guillaum and Limnocitrus littoralis (Miq.) Swingle
    Article Snippet: .. Viruses and Antiviral Assays All viruses were purchased from American Type Culture Collection (ATCC), with the exclusion of Human Immunodeficiency Virus type-1 (HIV-1) and Yellow Fever Virus (YFV): IIIB laboratory strain of HIV-1 was obtained from the supernatant of the acutely infected H9/IIIB cells (NIH 1983); yellow fever virus (YFV) (strain 17-D vaccine (Stamaril Pasteur J07B01)); bovine viral diarrhoea virus (BVDV) (strain NADL (ATCC VR-534)); coxsackie type B4 (CV-B4) (strain J.V.B. (ATCC VR-184)); human enterovirus C (poliovirus type-1 (Sb-1) (Sabin strain Chat (ATCC VR-1562)); vesicular stomatitis virus (VSV) (lab strain Indiana (ATCC VR 1540)); human respiratory syncytial virus (hRSV) (strain A2 (ATCC VR-1540)); reovirus type-1 (Reo-1) (simian virus 12, strain 3651 (ATCC VR- 214)), vaccinia virus (VV) (vaccine strain Elstree-Lister (ATCC VR-1549)); human herpes 1 (HSV-1) (strain KOS (ATCC VR-1493)).Viruses were maintained in our laboratory and propagated in suitable cell lines. ..

    Article Title: Dichloro-Phenyl-Benzotriazoles: A New Selective Class of Human Respiratory Syncytial Virus Entry Inhibitors
    Article Snippet: .. Viruses representative of positive-sense, single-stranded RNAs (ssRNA+) were: (i) Retroviridae : the IIIB laboratory strain of HIV-1, obtained from the supernatant of the persistently infected H9/IIIB cells (NIH 1983); (ii) Flaviviridae : Yellow Fever Virus (YFV) [strain 17-D vaccine (Stamaril Pasteur J07B01)] and Bovine Viral Diarrhea Virus (BVDV) [strain NADL (ATCC VR-534)]; (iii) Picornaviridae : Human Enterovirus B [coxsackie type B5 (CV-B5), strain Faulkner (ATCC VR-185)] and Human Enterovirus C [poliovirus type-1 (Sb-1), Sabin strain Chat (ATCC VR-1562)]. ..

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  • 94
    ATCC bvdv nadl strain
    Antibody neutralization against <t>BVDV</t> viral infection. (A) Normal MDBK cells grown in cell culture medium. (B) MDBK cells incubated with BVDV <t>NADL</t> virus pretreated with the serum sample (1:320 dilution) pooled from mice immunized with the FanC-STa-E2 fusion.
    Bvdv Nadl Strain, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bvdv nadl strain/product/ATCC
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    bvdv nadl strain - by Bioz Stars, 2021-09
    94/100 stars
      Buy from Supplier

    93
    ATCC bvdv strain nadl
    Fusion from without of <t>BVDV</t> in the presence or absence of DTT. MDBK cells were inoculated with BVDV strain <t>NADL</t> and briefly shifted to 37°C at the indicated pH in the presence or absence of 10 mM DTT; virus uptake via endocytosis was blocked by replacing buffer with DMEM containing bafilomycin A1. Higher concentrations of DTT could not be used due to high cell toxicity. The numbers of infectious centers were determined 12 to 16 h p.i. The columns represent mean values of triplicate experiments; bars indicate maximum and minimum values.
    Bvdv Strain Nadl, supplied by ATCC, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bvdv strain nadl/product/ATCC
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    bvdv strain nadl - by Bioz Stars, 2021-09
    93/100 stars
      Buy from Supplier

    Image Search Results


    Antibody neutralization against BVDV viral infection. (A) Normal MDBK cells grown in cell culture medium. (B) MDBK cells incubated with BVDV NADL virus pretreated with the serum sample (1:320 dilution) pooled from mice immunized with the FanC-STa-E2 fusion.

    Journal: Clinical and Vaccine Immunology : CVI

    Article Title: A Multiepitope Fusion Antigen Elicits Neutralizing Antibodies against Enterotoxigenic Escherichia coli and Homologous Bovine Viral Diarrhea Virus In Vitro

    doi: 10.1128/CVI.00249-13

    Figure Lengend Snippet: Antibody neutralization against BVDV viral infection. (A) Normal MDBK cells grown in cell culture medium. (B) MDBK cells incubated with BVDV NADL virus pretreated with the serum sample (1:320 dilution) pooled from mice immunized with the FanC-STa-E2 fusion.

    Article Snippet: The BVDV NADL strain (ATCC VR-534) was used to amplify the BVDV E2 gene and also as a challenge strain in viral neutralizing activity assays.

    Techniques: Neutralization, Infection, Cell Culture, Incubation, Mouse Assay

    Fusion from without of BVDV in the presence or absence of DTT. MDBK cells were inoculated with BVDV strain NADL and briefly shifted to 37°C at the indicated pH in the presence or absence of 10 mM DTT; virus uptake via endocytosis was blocked by replacing buffer with DMEM containing bafilomycin A1. Higher concentrations of DTT could not be used due to high cell toxicity. The numbers of infectious centers were determined 12 to 16 h p.i. The columns represent mean values of triplicate experiments; bars indicate maximum and minimum values.

    Journal: Journal of Virology

    Article Title: Acid-Resistant Bovine Pestivirus Requires Activation for pH-Triggered Fusion during Entry

    doi: 10.1128/JVI.79.7.4191-4200.2005

    Figure Lengend Snippet: Fusion from without of BVDV in the presence or absence of DTT. MDBK cells were inoculated with BVDV strain NADL and briefly shifted to 37°C at the indicated pH in the presence or absence of 10 mM DTT; virus uptake via endocytosis was blocked by replacing buffer with DMEM containing bafilomycin A1. Higher concentrations of DTT could not be used due to high cell toxicity. The numbers of infectious centers were determined 12 to 16 h p.i. The columns represent mean values of triplicate experiments; bars indicate maximum and minimum values.

    Article Snippet: A total of 5 × 105 MDBK cells or MDBK cells expressing either form of dynamin were grown in the absence or presence of 10 μg of doxycycline/ml for 24 h. A total of 2 × 105 PFU of BVDV strain NADL or SinV was adsorbed for 1 h at 4°C and washed with DMEM, followed by incubation at 37°C.

    Techniques:

    pH stability of BVDV. A total of 2 × 10 6 PFU of BVDV strain NADL were incubated in citrate-phosphate buffers of a defined pH (pH 3.0 to 7.0) in the presence of 10 mM DTT for 15 min at 25°C and titrated on MDBK cells. The same experiment was performed in the absence of DTT, but only the infectivity after treatment at pH 3.0 and 7.0 is indicated. The columns represent mean values of triplicate experiments; bars indicate maximum and minimum values.

    Journal: Journal of Virology

    Article Title: Acid-Resistant Bovine Pestivirus Requires Activation for pH-Triggered Fusion during Entry

    doi: 10.1128/JVI.79.7.4191-4200.2005

    Figure Lengend Snippet: pH stability of BVDV. A total of 2 × 10 6 PFU of BVDV strain NADL were incubated in citrate-phosphate buffers of a defined pH (pH 3.0 to 7.0) in the presence of 10 mM DTT for 15 min at 25°C and titrated on MDBK cells. The same experiment was performed in the absence of DTT, but only the infectivity after treatment at pH 3.0 and 7.0 is indicated. The columns represent mean values of triplicate experiments; bars indicate maximum and minimum values.

    Article Snippet: A total of 5 × 105 MDBK cells or MDBK cells expressing either form of dynamin were grown in the absence or presence of 10 μg of doxycycline/ml for 24 h. A total of 2 × 105 PFU of BVDV strain NADL or SinV was adsorbed for 1 h at 4°C and washed with DMEM, followed by incubation at 37°C.

    Techniques: Incubation, Infection

    Effect of chlorpromazine and β-MCD on BVDV and BHV-1 infection. BVDV NADL (dark gray bars) or BHV-1 (light gray bars) was adsorbed to MDBK cells, and the effects of chlorpromazine and β-MCD on infection were investigated. Susceptibility to BVDV infection was decreased up to 1,000-fold, whereas BHV-1 infection was inhibited five-fold by β-MCD but not by chlorpromazine. The columns represent mean values of duplicate experiments; bars indicate maximum and minimum values.

    Journal: Journal of Virology

    Article Title: Acid-Resistant Bovine Pestivirus Requires Activation for pH-Triggered Fusion during Entry

    doi: 10.1128/JVI.79.7.4191-4200.2005

    Figure Lengend Snippet: Effect of chlorpromazine and β-MCD on BVDV and BHV-1 infection. BVDV NADL (dark gray bars) or BHV-1 (light gray bars) was adsorbed to MDBK cells, and the effects of chlorpromazine and β-MCD on infection were investigated. Susceptibility to BVDV infection was decreased up to 1,000-fold, whereas BHV-1 infection was inhibited five-fold by β-MCD but not by chlorpromazine. The columns represent mean values of duplicate experiments; bars indicate maximum and minimum values.

    Article Snippet: A total of 5 × 105 MDBK cells or MDBK cells expressing either form of dynamin were grown in the absence or presence of 10 μg of doxycycline/ml for 24 h. A total of 2 × 105 PFU of BVDV strain NADL or SinV was adsorbed for 1 h at 4°C and washed with DMEM, followed by incubation at 37°C.

    Techniques: Infection

    Expression of Dyn K44A reduces susceptibility to BVDV infection. (a) Immunoblot of MDBK Tet on dynamin-overexpressing cell lines. Crude cell lysates from equal numbers of cells grown in the presence or absence of 10 μg of doxycycline/ml were separated. After induction, a 99-kDa band of each HA-tagged protein is visible. (b) Inhibition of BVDV NADL/SinV infection by overexpression of dominant-negative Dyn K44A . Each indicated cell line was tested for its susceptibility to BVDV or SinV infection by inoculation with 2 × 10 5 PFU of BVDV strain NADL or SinV, respectively. MDBK cells overexpressing mutant dynamin after induction with doxycycline exhibited a 10-fold- reduced susceptibility compared to MDBK cells. The columns represent mean values of triplicate experiments; bars indicate maximum and minimum values.

    Journal: Journal of Virology

    Article Title: Acid-Resistant Bovine Pestivirus Requires Activation for pH-Triggered Fusion during Entry

    doi: 10.1128/JVI.79.7.4191-4200.2005

    Figure Lengend Snippet: Expression of Dyn K44A reduces susceptibility to BVDV infection. (a) Immunoblot of MDBK Tet on dynamin-overexpressing cell lines. Crude cell lysates from equal numbers of cells grown in the presence or absence of 10 μg of doxycycline/ml were separated. After induction, a 99-kDa band of each HA-tagged protein is visible. (b) Inhibition of BVDV NADL/SinV infection by overexpression of dominant-negative Dyn K44A . Each indicated cell line was tested for its susceptibility to BVDV or SinV infection by inoculation with 2 × 10 5 PFU of BVDV strain NADL or SinV, respectively. MDBK cells overexpressing mutant dynamin after induction with doxycycline exhibited a 10-fold- reduced susceptibility compared to MDBK cells. The columns represent mean values of triplicate experiments; bars indicate maximum and minimum values.

    Article Snippet: A total of 5 × 105 MDBK cells or MDBK cells expressing either form of dynamin were grown in the absence or presence of 10 μg of doxycycline/ml for 24 h. A total of 2 × 105 PFU of BVDV strain NADL or SinV was adsorbed for 1 h at 4°C and washed with DMEM, followed by incubation at 37°C.

    Techniques: Expressing, Infection, Inhibition, Over Expression, Dominant Negative Mutation, Mutagenesis

    Effect of different inhibitors of endosomal acidification on BVDV and BHV-1 infection. Directly after adsorption of BVDV NADL (dark gray bars) or BHV-1 (light gray bars) to MDBK cells, different inhibitors of endosomal acidification (bafilomycin A1, chloroquine, or ammonium chloride) were applied to determine the pH dependence of viral entry. Each inhibitor of endosomal acidification blocks BVDV infection in a concentration-dependent manner, whereas BHV-1 infection is not affected. The columns represent mean values of duplicate experiments; bars indicate maximum and minimum values.

    Journal: Journal of Virology

    Article Title: Acid-Resistant Bovine Pestivirus Requires Activation for pH-Triggered Fusion during Entry

    doi: 10.1128/JVI.79.7.4191-4200.2005

    Figure Lengend Snippet: Effect of different inhibitors of endosomal acidification on BVDV and BHV-1 infection. Directly after adsorption of BVDV NADL (dark gray bars) or BHV-1 (light gray bars) to MDBK cells, different inhibitors of endosomal acidification (bafilomycin A1, chloroquine, or ammonium chloride) were applied to determine the pH dependence of viral entry. Each inhibitor of endosomal acidification blocks BVDV infection in a concentration-dependent manner, whereas BHV-1 infection is not affected. The columns represent mean values of duplicate experiments; bars indicate maximum and minimum values.

    Article Snippet: A total of 5 × 105 MDBK cells or MDBK cells expressing either form of dynamin were grown in the absence or presence of 10 μg of doxycycline/ml for 24 h. A total of 2 × 105 PFU of BVDV strain NADL or SinV was adsorbed for 1 h at 4°C and washed with DMEM, followed by incubation at 37°C.

    Techniques: Infection, Adsorption, Concentration Assay

    Fusion from without of BVDV and SinV. MDBK cells were inoculated with BVDV strain NADL or SinV, respectively, for 1 h at 4°C. Medium was replaced by prewarmed buffers of the indicated pH, followed by incubation for 2 min at 37°C. Viral uptake via endocytosis was blocked by replacing buffer with DMEM containing bafilomycin A1. Since fusion from without is cell type specific, SinV was used as control. The columns represent mean values of triplicate experiments; bars indicate maximum and minimum values.

    Journal: Journal of Virology

    Article Title: Acid-Resistant Bovine Pestivirus Requires Activation for pH-Triggered Fusion during Entry

    doi: 10.1128/JVI.79.7.4191-4200.2005

    Figure Lengend Snippet: Fusion from without of BVDV and SinV. MDBK cells were inoculated with BVDV strain NADL or SinV, respectively, for 1 h at 4°C. Medium was replaced by prewarmed buffers of the indicated pH, followed by incubation for 2 min at 37°C. Viral uptake via endocytosis was blocked by replacing buffer with DMEM containing bafilomycin A1. Since fusion from without is cell type specific, SinV was used as control. The columns represent mean values of triplicate experiments; bars indicate maximum and minimum values.

    Article Snippet: A total of 5 × 105 MDBK cells or MDBK cells expressing either form of dynamin were grown in the absence or presence of 10 μg of doxycycline/ml for 24 h. A total of 2 × 105 PFU of BVDV strain NADL or SinV was adsorbed for 1 h at 4°C and washed with DMEM, followed by incubation at 37°C.

    Techniques: Incubation

    Comparison of CxxC motifs from different viruses. A decapeptide of BVDV strain NADL and CSFV Alfort E2 containing the CxxC motif was aligned to the corresponding sequence in rubella virus E1 (GI:33415288) and Mo-MuLV gPr80 glycosylated envelope polyprotein (GI:18448745). The numbers denote the position of the decapeptide in the respective protein; the conserved CxxC motif is indicated above the sequence. Conserved cysteine residues are boxed.

    Journal: Journal of Virology

    Article Title: Acid-Resistant Bovine Pestivirus Requires Activation for pH-Triggered Fusion during Entry

    doi: 10.1128/JVI.79.7.4191-4200.2005

    Figure Lengend Snippet: Comparison of CxxC motifs from different viruses. A decapeptide of BVDV strain NADL and CSFV Alfort E2 containing the CxxC motif was aligned to the corresponding sequence in rubella virus E1 (GI:33415288) and Mo-MuLV gPr80 glycosylated envelope polyprotein (GI:18448745). The numbers denote the position of the decapeptide in the respective protein; the conserved CxxC motif is indicated above the sequence. Conserved cysteine residues are boxed.

    Article Snippet: A total of 5 × 105 MDBK cells or MDBK cells expressing either form of dynamin were grown in the absence or presence of 10 μg of doxycycline/ml for 24 h. A total of 2 × 105 PFU of BVDV strain NADL or SinV was adsorbed for 1 h at 4°C and washed with DMEM, followed by incubation at 37°C.

    Techniques: Sequencing

    The NS5B C terminus conditions the production of infectious viral progeny. (A) Infectivity of in vitro-transcribed RNA from WT BVDV and NS5B mutant genomes determined by infectious center assay. BHK-21 cells were electroporated with RNA transcripts from pNADLp15 or from p5Bgdd, p5B-741, or p5B-L726P serially diluted and plated with additional fresh MDBK cells with agar overlay. (B) Immunofluorescence assay of cells electroporated with in vitro-transcribed RNA. RNA transcripts (2 μg) from 5Bgdd (1), mutant 5B-741 (3), 5B-L726P (5), N-ΔINS ncBVDV (7), and WT BVDV NADLp15 (9) were electroporated into RD420 cells and RNA replication was monitored by IIF using anti-E2 antibodies. Release of infectious virus was assessed by harvesting the culture medium (panels 1, 3, 5, 7, and 9) at 24 hpe, inoculating into fresh RD420 cells, and monitoring by IIF using anti-E2 antibodies (panels 2, 4, 6, 8, and 10, respectively). (C) Northern blot analysis of total RNA from cells electroporated with RNA from NS5B mutants. (a) Total RNA from BHK-21 cells electroporated with 5Bgdd (1), 5B-741 (2), 5B-L726P (3), N-ΔINS (4), and N-p15 (5) was analyzed, and 28S rRNA is indicated as a control. (b) The transferred RNA was hybridized to a 32 ). (D) Western blot analysis to demonstrate expression of structural and NS proteins in electroporated cells with mutant NS5B RNA. Panels: 1, 5Bgdd; 2, 5B741; 3, L726P; 4, N-ΔINS; 5, NADLp15. The total protein was resolved by SDS-polyacrylamide gel electrophoresis and probed with anti-E2 and anti-NS3 MAbs, and signal was developed with a chemiluminescence detection system. Molecular mass markers are indicated (in kilodaltons). (E) Plaque phenotype of NS5B mutants. Plaques formed by revertant i-5B-741rev (a), the virus recovered from the engineered mutant i-5B-L726P (b), or WT BVDV strain NADL (c) on bovine cells infected for 60 h. The average diameters of the plaques are indicated.

    Journal: Journal of Virology

    Article Title: Involvement of a Bovine Viral Diarrhea Virus NS5B Locus in Virion Assembly †

    doi: 10.1128/JVI.78.18.9612-9623.2004

    Figure Lengend Snippet: The NS5B C terminus conditions the production of infectious viral progeny. (A) Infectivity of in vitro-transcribed RNA from WT BVDV and NS5B mutant genomes determined by infectious center assay. BHK-21 cells were electroporated with RNA transcripts from pNADLp15 or from p5Bgdd, p5B-741, or p5B-L726P serially diluted and plated with additional fresh MDBK cells with agar overlay. (B) Immunofluorescence assay of cells electroporated with in vitro-transcribed RNA. RNA transcripts (2 μg) from 5Bgdd (1), mutant 5B-741 (3), 5B-L726P (5), N-ΔINS ncBVDV (7), and WT BVDV NADLp15 (9) were electroporated into RD420 cells and RNA replication was monitored by IIF using anti-E2 antibodies. Release of infectious virus was assessed by harvesting the culture medium (panels 1, 3, 5, 7, and 9) at 24 hpe, inoculating into fresh RD420 cells, and monitoring by IIF using anti-E2 antibodies (panels 2, 4, 6, 8, and 10, respectively). (C) Northern blot analysis of total RNA from cells electroporated with RNA from NS5B mutants. (a) Total RNA from BHK-21 cells electroporated with 5Bgdd (1), 5B-741 (2), 5B-L726P (3), N-ΔINS (4), and N-p15 (5) was analyzed, and 28S rRNA is indicated as a control. (b) The transferred RNA was hybridized to a 32 ). (D) Western blot analysis to demonstrate expression of structural and NS proteins in electroporated cells with mutant NS5B RNA. Panels: 1, 5Bgdd; 2, 5B741; 3, L726P; 4, N-ΔINS; 5, NADLp15. The total protein was resolved by SDS-polyacrylamide gel electrophoresis and probed with anti-E2 and anti-NS3 MAbs, and signal was developed with a chemiluminescence detection system. Molecular mass markers are indicated (in kilodaltons). (E) Plaque phenotype of NS5B mutants. Plaques formed by revertant i-5B-741rev (a), the virus recovered from the engineered mutant i-5B-L726P (b), or WT BVDV strain NADL (c) on bovine cells infected for 60 h. The average diameters of the plaques are indicated.

    Article Snippet: The cytopathogenic (cp) BVDV strain NADL was obtained from the American Type Culture Collection ( ).

    Techniques: Infection, In Vitro, Mutagenesis, Immunofluorescence, Northern Blot, Western Blot, Expressing, Polyacrylamide Gel Electrophoresis