Journal: Viruses

Article Title: A Novel Hemocyte-Specific Small Protein Participates in White Spot Syndrome Virus Infection via Binding to Viral Envelope Protein

doi: 10.3390/v15010227

Figure Lengend Snippet: Encoding capability of start codon in the predicted ORF in LvHSSP transcript. ( A ) Diagram of the GFP fusion constructs used for transfection. The start codon ATGGTG of the GFP (GFPwt) gene is mutated to ATTGTT (GFPmut). The start codon ATG of the ORF in LvHSSP transcript is mutated to ATT. ( B ) GFP fluorescence signals in different constructs of transfected Sf9 cells.

Article Snippet: Sf9 cells were purchased from the China Center for Type Culture Collection (Wuhan, China), cultured in Sf-900TM II SFM (Gibco, Grand Island, NE, USA) at 27 °C, and sub-cultured every 3–4 days.

Techniques: Construct, Transfection, Fluorescence

Journal: Viruses

Article Title: A Lac Repressor-Inducible Baculovirus Expression Vector for Controlling Adeno-Associated Virus Capsid Ratios

doi: 10.3390/v16010051

Figure Lengend Snippet: An Original and Commonly Used rBEV (Bacmid) RepCap Dual Cassette Design for Producing rAAV Sf9 Cells. The overlapping Rep78/Rep52 and Cap-related VP1/VP2/VP3/AAP genes are shown in context with the bipartite baculovirus very late polh / p10 promoter driving expression of Rep and Cap. This 4389 bp dual cassette is cloned into a single location in the BEV. Non-canonical translation start-site sequences are indicated. Rep genes Rep78 (1866 bp) and Rep52 (1194 bp) have common ORFs and 3′ regions. Cap genes VP1 (2111 bp), VP2 (1800 bp), and VP3 (1605 bp) have common ORF 3′ regions. The AAP gene ORF is out of frame from Cap genes, begins 526 bp from the VP1 translational initiation site, and is 594 bp. Only the VP1 and VP2 ORFs overlap the AAP ORF.

Article Snippet: Spodoptera frugiperda ( Sf9 ) cells used were a Voyager Therapeutics clonal isolate of the Sf9 cell line [ ], which was a clonal derivative of the IPLB-SF-21-AE cell line [ ].

Techniques: Expressing, Clone Assay

Journal: Viruses

Article Title: A Lac Repressor-Inducible Baculovirus Expression Vector for Controlling Adeno-Associated Virus Capsid Ratios

doi: 10.3390/v16010051

Figure Lengend Snippet: Testing LacR Regulation of LacOVP1 Expression in the Context of Baculovirus Infection. Bacmid VB-LacR-LacOVP1 is schematically illustrated ( A ). The location and transcription directions of baculovirus genes gp64 , gta , and Lef1 are shown. This bacmid included a LacO-p10-LacO promoter VP1 cassette ( B ) in the VC locus and a polh promoter LacR cassette in the egt locus ( C ). Sf9 cells were infected at 10 MOI in the presence of different concentrations of IPTG. At 72 hpi, infected Sf9 cells were collected, fractionated by SDS-PAGE, and then Western blot probed with an anti-capsid monoclonal antibody ( D ) and an anti-lac repressor antibody ( E ). Relative abundances of ECL-detected proteins were quantified using ImageJ software (version 1.53) and plotted ( F ).

Article Snippet: Spodoptera frugiperda ( Sf9 ) cells used were a Voyager Therapeutics clonal isolate of the Sf9 cell line [ ], which was a clonal derivative of the IPLB-SF-21-AE cell line [ ].

Techniques: Expressing, Infection, SDS Page, Western Blot, Software

Journal: Viruses

Article Title: A Lac Repressor-Inducible Baculovirus Expression Vector for Controlling Adeno-Associated Virus Capsid Ratios

doi: 10.3390/v16010051

Figure Lengend Snippet: Testing bacmid VB-LacR-Rep-VP3 and Validating a Hybrid Early/Late/Very Late Promoter for LacR Expression. The bacmid VB-LacR-Rep-VP3 is schematically illustrated ( A ). This bacmid was based on VoyBac1.1 and had a LacR-Rep-VP3 multi-cassette inserted into the Tn7 locus. The Rep and VP3 were in opposing orientations and had polh and p10 promoters, respectively ( B ). The LacR ORF was downstream of Rep and in the same orientation. The original promoter design for LacR in this multi-cassette was a hybrid Opgp64 - polh promoter ( C ). This promoter had two late ATAAG transcriptional initiation sites: an early CAGT transcriptional initiation site in the Opgp64 promoter and a very late ATAAG transcriptional initiation site in the polh promoter. A minicistron was present in the polh promoter upstream of the polh ATAAG transcriptional initiation site ( D ). A mutagenic PCR primer was designed to remove two ATG translational initiation sites in the minicistron ( B ). A mutagenic PCR primer was used to remove the minicistron translational initiation sites from the polh promoter, resulting in an Opgp64polh mt LacR cassette ( E ). Two versions of VB-LacR-Rep-VP3 were made: Opgp64polh wt LacR-Rep-VP3 (original version) and Opgp64polh mt LacR-Rep-VP3 (minicistron mutated version). Sf9 cells were infected at 10 MOI with the two versions of bacmid VB-LacR-Rep-VP3, and insect cells were collected at different hours post-infection. Cell proteins were fractionated by SDS-PAGE, and then Western blots were probed with anti-Cap or anti-LacR antibodies. The relative abundances of Western-detected VP3 ( F ) and LacR ( G ) are plotted on the graphs.

Article Snippet: Spodoptera frugiperda ( Sf9 ) cells used were a Voyager Therapeutics clonal isolate of the Sf9 cell line [ ], which was a clonal derivative of the IPLB-SF-21-AE cell line [ ].

Techniques: Expressing, Infection, SDS Page, Western Blot

Journal: Viruses

Article Title: A Lac Repressor-Inducible Baculovirus Expression Vector for Controlling Adeno-Associated Virus Capsid Ratios

doi: 10.3390/v16010051

Figure Lengend Snippet: Bacmid VB-LacRepCap and Time Course of Cap and Lac Expression in Sf9 Cells. The bacmid VB-LacRepCap is illustrated schematically ( A ). This bacmid was a combination of the bacmids VB-LacR-VP1 and VB-LacR-Rep-VP3. VB-LacRepCap also included a new LacOVP2 cassette ( B ) in the gta locus. Sf9 cells were infected at 10 MOI with VB-LacRepCap. Cells were collected at different hours post-infection, from 0 hpi to 50 hpi. Cell proteins were fractionated by SDS-PAGE and then Western blot probed with anti-capsid monoclonal antibody and anti-lac repressor antibody ( C ). ( D ) ECL signals of Western-detected proteins were quantified using ImageJ software. ( E ) Relative capsid VP ratios are expressed in bar graph format.

Article Snippet: Spodoptera frugiperda ( Sf9 ) cells used were a Voyager Therapeutics clonal isolate of the Sf9 cell line [ ], which was a clonal derivative of the IPLB-SF-21-AE cell line [ ].

Techniques: Expressing, Infection, SDS Page, Western Blot, Software

Journal: Viruses

Article Title: A Lac Repressor-Inducible Baculovirus Expression Vector for Controlling Adeno-Associated Virus Capsid Ratios

doi: 10.3390/v16010051

Figure Lengend Snippet: Titration of IPTG Induction of VP1 and VP2 expression. Sf9 cells were infected at 10 MOI with VB-LacRepCap bacmid in the presence of different concentrations of IPTG. Cells were collected at 72 hpi, and their proteins were fractionated by SDS-PAGE and Western blot probed with an anti-Capsid monoclonal antibody. Relative capsid abundances in infected cells were observed by Western blot ( A ) and ECL signals, which were quantified using ImageJ software. Relative VP1, VP2, and VP3 capsid abundances in infected cells are graphed relative to IPTG concentration ( B ). Calculated capsid ratios are expressed in bar graph format ( C ).

Article Snippet: Spodoptera frugiperda ( Sf9 ) cells used were a Voyager Therapeutics clonal isolate of the Sf9 cell line [ ], which was a clonal derivative of the IPLB-SF-21-AE cell line [ ].

Techniques: Titration, Expressing, Infection, SDS Page, Western Blot, Software, Concentration Assay

Journal: Viruses

Article Title: A Lac Repressor-Inducible Baculovirus Expression Vector for Controlling Adeno-Associated Virus Capsid Ratios

doi: 10.3390/v16010051

Figure Lengend Snippet: Optimizing VB-ITR-SEAP and VB-LacRepCap Bacmid Co-Infection Ratios. Sf9 cells were co-infected with VB-ITR-SEAP and VB-LacRepCap bacmids at different co-infection ratios at 0.1 MOI in the presence of 0 μM and 200 μM IPTG. A 72 h post-infection, ( A ) Cell lysate protein samples were fractionated by SDS-PAGE and Western blot probed with anti-capsid antibody. ( B ) Crude cell lysate titers for ITR-SEAP-ITR transgenes were determined by Q-PCR. ( C ) Corresponding estimated capsid ratios are expressed in bar graph format for 0 μM IPTG and 200 μM IPTG.

Article Snippet: Spodoptera frugiperda ( Sf9 ) cells used were a Voyager Therapeutics clonal isolate of the Sf9 cell line [ ], which was a clonal derivative of the IPLB-SF-21-AE cell line [ ].

Techniques: Infection, SDS Page, Western Blot

Journal: Viruses

Article Title: A Lac Repressor-Inducible Baculovirus Expression Vector for Controlling Adeno-Associated Virus Capsid Ratios

doi: 10.3390/v16010051

Figure Lengend Snippet: Effect of IPTG on total AAV Capsid Protein Abundance in Cells and in Purified AAV Capsids. Sf9 cells were co-infected with VB-ITR-SEAP and VB-LacRepCap bacmids at 0.1 MOI and co-infection ratios of 1:1, 1:3, and 1:6 in the presence of different concentrations of IPTG. At 72 h post-infection, protein samples were obtained from cell lysates ( A ) and from sucrose cushion ultracentrifugation pellet fractions of cell lysates ( B ). All samples were fractionated by SDS-PAGE, and Western blots were probed with an anti-capsid monoclonal antibody. Immunolabeled capsid proteins were detected by ECL and quantified using ImageJ software. Total capsid protein abundances in cell lysates ( C ) and sucrose cushion pellets ( D ) were plotted relative to IPTG concentration for each co-infection group. Total capsid abundances at different co-infection ratios are represented on graphs as open triangles (1:1 ratio), black diamonds (1:3 ratio), and open circles (1:6 ratio). The estimated capsid ratios for the 1:6 co-infection are expressed in bar graph format for cell lysate ( E ) and sucrose cushion samples ( F ).

Article Snippet: Spodoptera frugiperda ( Sf9 ) cells used were a Voyager Therapeutics clonal isolate of the Sf9 cell line [ ], which was a clonal derivative of the IPLB-SF-21-AE cell line [ ].

Techniques: Quantitative Proteomics, Purification, Infection, SDS Page, Western Blot, Immunolabeling, Software, Concentration Assay

Journal: Viruses

Article Title: A Lac Repressor-Inducible Baculovirus Expression Vector for Controlling Adeno-Associated Virus Capsid Ratios

doi: 10.3390/v16010051

Figure Lengend Snippet: Titer and Potency of AAV Produced at Different IPTG Concentrations. Sf9 cells were co-infected with VB-ITR-SEAP and VB-LacRepCap bacmids at 0.1 MOI. At 72 h post-infection, lysates were made. Sucrose cushion-purified AAV sample Q-PCR titers are plotted against IPTG concentration ( A ). Titers generated at ITR-SEAP-ITR:LacRepCap co-infection ratios of 1:1 (open triangle), 1:3 (black diamond), and 1:6 (open circle) are shown. Cells were lysed, and AAV capsids were purified by sucrose cushion ultracentrifugation. These AAV samples were used to transduce HEK 293T cells, and the alkaline phosphatase activity was measured after 4 days. Potency was measured as alkaline phosphatase activity relative to genome titer, which is plotted against IPTG concentration ( B ). The graph is split-scaled to emphasize the potency values of capsids produced at IPTG concentrations between 0 μM and 10 μM.

Article Snippet: Spodoptera frugiperda ( Sf9 ) cells used were a Voyager Therapeutics clonal isolate of the Sf9 cell line [ ], which was a clonal derivative of the IPLB-SF-21-AE cell line [ ].

Techniques: Produced, Infection, Purification, Concentration Assay, Generated, Transduction, Activity Assay

Journal: Viruses

Article Title: A Lac Repressor-Inducible Baculovirus Expression Vector for Controlling Adeno-Associated Virus Capsid Ratios

doi: 10.3390/v16010051

Figure Lengend Snippet: Comparison of Capsid Ratios Before and After Affinity Purification. Sf9 cells were cultured at an 800 mL scale and co-infected at 0.001 MOI with VB-ITR-SEAP and VB-LacRepCap bacmids at a 1:6 co-infection ratio in the presence of 0 μM, 2 μM, 10 μM, and 50 μM IPTG. At 72 h post-infection, cells were lysed and capsids were affinity purified, followed by further sucrose cushion purification. Total capsid proteins in cell lysates were analyzed by Western blot and quantified from ECL images using ImageJ software ( A ). Affinity purified capsids ( B ) and affinity/sucrose cushion purified capsids ( C ) were analyzed by CE-SDS.

Article Snippet: Spodoptera frugiperda ( Sf9 ) cells used were a Voyager Therapeutics clonal isolate of the Sf9 cell line [ ], which was a clonal derivative of the IPLB-SF-21-AE cell line [ ].

Techniques: Comparison, Affinity Purification, Cell Culture, Infection, Purification, Western Blot, Software

Journal: Viruses

Article Title: A Lac Repressor-Inducible Baculovirus Expression Vector for Controlling Adeno-Associated Virus Capsid Ratios

doi: 10.3390/v16010051

Figure Lengend Snippet: Potency and Percent Full of Affinity purified and affinity/sucrose cushion purified AAV capsids. Purified rAAV from VB-ITR-SEAP and VB-LacRepCap bacmid co-infection of Sf9 cells at an 800 mL scale was used to transduce HEK 293T cells. The alkaline phosphatase activity was measured after 4 days. Potency was measured as alkaline phosphatase activity relative to AAV viral genome (vg) added to HEK 293T cells and is plotted against IPTG concentration for affinity purified AAV ( A ) and for affinity/sucrose cushion purified AAV ( B ). Affinity purified capsids were also subjected to SEC-MALS analysis, and the percent full (ITR-SEAP-ITR transgene genomes) in capsids relative to IPTG concentration is shown ( C ).

Article Snippet: Spodoptera frugiperda ( Sf9 ) cells used were a Voyager Therapeutics clonal isolate of the Sf9 cell line [ ], which was a clonal derivative of the IPLB-SF-21-AE cell line [ ].

Techniques: Affinity Purification, Purification, Infection, Transduction, Activity Assay, Concentration Assay

Journal: Journal of Biomedical Science

Article Title: Hepatitis B virus virion secretion is a CRM1-spike-mediated late event

doi: 10.1186/s12929-022-00827-w

Figure Lengend Snippet: Lack of HBc secretion of an HBc double mutant L65A/V89A in human hepatocytes and insect cells. A A cartoon illustration of secreted HBV particles, including virions, naked capsids, and HBsAg particles. Left panel: genome-containing and genome-free empty virions contain both surface antigen and core proteins; middle panel: genome-containing and genome-free naked capsids contain no surface antigens; right panel: HBV surface antigen particles contain no core protein. B Upper panel: Two CRM1-dependent nuclear export signals (HBc NES CRM1 -I and NES CRM1 -II) were mapped to the tip of the spike of HBc particles . The double mutations L65A/V89A at NES I + II are shown in red color. Lower panel: No secreted naked capsids were detected by ELISA in the media of Sf9 insect cells transfected with an HBc mutant L65A/V89A expression vector (left panel). Nor can any naked capsids be detected in the media of HuH-7 human hepatocytes transfected with an HBV replicon (right panel). The ELISA values were calculated from three independent repeat experiments. WT: wild type HBV. Mock: un-transfected cells

Article Snippet: Insect cells Sf9 ( Spodoptera frugiperda 9 ) were purchased from BCRC (Bioresource Collection and Research Center, Taiwan; ATCC number: CRL-1711).

Techniques: Mutagenesis, Enzyme-linked Immunosorbent Assay, Transfection, Expressing, Plasmid Preparation

Journal: PLoS ONE

Article Title: The FP25K Acts as a Negative Factor for the Infectivity of AcMNPV Budded Virus

doi: 10.1371/journal.pone.0128471

Figure Lengend Snippet: (A) Construction of fp25k knockout bacmid. The fp25k gene in AcBacΔ cc bacmid was deleted and replaced with the zeocin resistance gene ( zeocin r ) through homologous recombination. (B) Strategy for construction of recombinant viruses, vAcΔ cc , vAcΔ cc Δ fp25k and vAcΔ cc Δ fp25k-rfp25k , the e gfp gene was inserted into the polyhedrin locus by transposition. (C) Transfection-infection assay of recombinant bacmids for viral propagation. At 48 h p.t., GFP-expressing cells were observed by fluorescence microscopy (upper panel). The fluorescent signals of GFP in cells infected by transfection supernatant were visualized at 72 h p.i. (lower panel). (D) One step growth curves of vAcΔ cc , vAcΔ cc Δ fp25k and vAcΔ cc Δ fp25k-rfp25k . Sf9 cells were infected with each virus at an MOI of 5, the supernatants were harvested at 0, 24, 48 and 96 h p.i. and determined for the production of infectious virus by EPDA. The results were transformed logarithmically. Each point represents the average titer from three independent infections. Error bars represent standard deviations.

Article Snippet: The Spodoptera frugiperda (Sf9) cell line [ ] (as gift from Prof. Just M. Vlak, Wageningen University, The Netherlands) was cultured in Grace’s insect medium (pH 6.0; Gibco-BRL), supplemented with 10% fetal bovine serum (FBS; Gibco-BRL) at 28°C.

Techniques: Knock-Out, Homologous Recombination, Recombinant, Transfection, Infection, Expressing, Fluorescence, Microscopy, Virus, Transformation Assay

Journal: PLoS ONE

Article Title: The FP25K Acts as a Negative Factor for the Infectivity of AcMNPV Budded Virus

doi: 10.1371/journal.pone.0128471

Figure Lengend Snippet: Sf9 cells were infected with vAcΔ cc or vAcΔ cc Δ fp25k at an MOI of 5. The supernatants from infected cells were collected at 0, 18, 24, 48, 72 and 96 h. p.i. Virus titers were determined by EPDA (A), genomic DNA was detected by qPCR (B), and the results were transformed logarithmically. Each point represents the average titer from three independent infections. Error bars represent standard deviations.

Article Snippet: The Spodoptera frugiperda (Sf9) cell line [ ] (as gift from Prof. Just M. Vlak, Wageningen University, The Netherlands) was cultured in Grace’s insect medium (pH 6.0; Gibco-BRL), supplemented with 10% fetal bovine serum (FBS; Gibco-BRL) at 28°C.

Techniques: Infection, Virus, Transformation Assay

Journal: PLoS ONE

Article Title: The FP25K Acts as a Negative Factor for the Infectivity of AcMNPV Budded Virus

doi: 10.1371/journal.pone.0128471

Figure Lengend Snippet: (A) Electron microscopy analysis. Sf9 cells were infected with vAcΔ cc or vAcΔ cc Δ fp25k at an MOI of 5. Infected cells were fixed with glutaraldehyde at 48, 64, 72 and 96 h p.i., and nucleocapsids occlusion was observed (i-viii). Enlargements of the blocked area in (a-h). Bars represent 1 μm and 250 nm. (B) Cellular viral particle determination. Sf9 cells were infected with vAcΔ cc or vAcΔ cc Δ fp25k (MOI = 5), total cellular DNA were collected at 0, 72 and 96 h p.i. and subjected to qPCR to determine the viral genome copy number. The data are from three independent experiments. Error bars represent standard deviation. Data were analyzed by two-tailed Student's t -test.

Article Snippet: The Spodoptera frugiperda (Sf9) cell line [ ] (as gift from Prof. Just M. Vlak, Wageningen University, The Netherlands) was cultured in Grace’s insect medium (pH 6.0; Gibco-BRL), supplemented with 10% fetal bovine serum (FBS; Gibco-BRL) at 28°C.

Techniques: Electron Microscopy, Infection, Standard Deviation, Two Tailed Test

Journal: PLoS ONE

Article Title: The FP25K Acts as a Negative Factor for the Infectivity of AcMNPV Budded Virus

doi: 10.1371/journal.pone.0128471

Figure Lengend Snippet: (A) Sf9 cells were infected with vAcΔ cc or vAcΔ cc Δ fp25k at an MOI of 5. Infected cells were collected at 48 h p.i. and analyzed by western blotting using the corresponding antibodies, the results were repeated twice. (B) Sf9 cells infected with vAcΔ cc or vAcΔ cc Δ fp25k (MOI = 5) were collected at 48 h p.i. and analyzed by qRT-PCR. The transcriptional levels of viral genes were normalized to the internal control 28S rRNA, and the transcription difference between vAcΔ cc - and vAcΔ cc Δ fp25k -infected cells was analyzed by the 2 -ΔΔC T method. The results of corresponding genes in vAcΔ cc -infected cells were set as 100%. The data are from three independent experiments. Error bars represent standard deviation. Data were analyzed by two-tailed Student's t -test. * P <0.05.

Article Snippet: The Spodoptera frugiperda (Sf9) cell line [ ] (as gift from Prof. Just M. Vlak, Wageningen University, The Netherlands) was cultured in Grace’s insect medium (pH 6.0; Gibco-BRL), supplemented with 10% fetal bovine serum (FBS; Gibco-BRL) at 28°C.

Techniques: Infection, Western Blot, Quantitative RT-PCR, Control, Standard Deviation, Two Tailed Test

Journal: PLoS ONE

Article Title: The FP25K Acts as a Negative Factor for the Infectivity of AcMNPV Budded Virus

doi: 10.1371/journal.pone.0128471

Figure Lengend Snippet: (A) Western blot analysis of structural proteins incorporated into recombinant BVs. Equal copy numbers of vAcΔ cc and vAcΔ cc Δ fp25k virions were purified from the supernatants of infected Sf9 cells at 48 h p.i. and subjected to SDS-PAGE, then blotted with antibodies against GP64, Ac23, and VP39. (B) Transformation assay. Genomic DNA isolated from vAcΔ cc or vAcΔ cc Δ fp25k BVs harvested at 36 h and 48 h p.i. was quantified by qPCR. Equal copy numbers (5×10 8 ) of genomic DNA was used to transform competent E . coli DH10B cells, and the number of CFUs was calculated. (C) Transfection assay. Equal copy numbers of genomic DNA isolated from vAcΔ cc and vAcΔ cc Δ fp25k BVs were transfected into Sf9 cell. Transfection efficiency was calculated as the number of cells expressing EGFP. The mean number of cells transfected by BV genomic DNA of control virus was set as 100%. Data are representative of three independent experiments. Error bars represent standard deviation. Data were analyzed by two-tailed Student’s t -test. * P <0.05.

Article Snippet: The Spodoptera frugiperda (Sf9) cell line [ ] (as gift from Prof. Just M. Vlak, Wageningen University, The Netherlands) was cultured in Grace’s insect medium (pH 6.0; Gibco-BRL), supplemented with 10% fetal bovine serum (FBS; Gibco-BRL) at 28°C.

Techniques: Western Blot, Recombinant, Purification, Infection, SDS Page, Transformation Assay, Isolation, Transfection, Expressing, Control, Virus, Standard Deviation, Two Tailed Test

Journal: PLoS ONE

Article Title: The FP25K Acts as a Negative Factor for the Infectivity of AcMNPV Budded Virus

doi: 10.1371/journal.pone.0128471

Figure Lengend Snippet: (A) Genomic organization of AcBacΔ cc - gp64 . (B) Western blot analysis of cells infected with vAcΔ cc or vAcΔ cc - gp64 and virions of vAcΔ cc or vAcΔ cc - gp64 . For expression of GP64 in infected cells, Sf9 cells were infected with vAcΔ cc or vAcΔ cc - gp64 at an MOI of 5. Infected cells were collected at 48 h p.i. and analyzed by western blotting. For GP64 incorporation into virions, equal copy numbers of vAcΔ cc and vAcΔ cc - gp64 virions were purified from the supernatants of infected Sf9 cells at 48 h p.i. and subjected to SDS-PAGE, then blotted with antibody against GP64, experiments were repeated twice. (C) One-step growth curve of vAcΔ cc and vAcΔ cc - gp64 , virus titers were determined by EPDA. Data are representative of three independent experiments. Error bars represent standard deviation.

Article Snippet: The Spodoptera frugiperda (Sf9) cell line [ ] (as gift from Prof. Just M. Vlak, Wageningen University, The Netherlands) was cultured in Grace’s insect medium (pH 6.0; Gibco-BRL), supplemented with 10% fetal bovine serum (FBS; Gibco-BRL) at 28°C.

Techniques: Western Blot, Infection, Expressing, Purification, SDS Page, Virus, Standard Deviation