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  • 99
    Thermo Fisher pvax 1 vector
    Agarose gel electrophoresis for ( A ) polymerase chain reaction (PCR) amplification of ROP8 gene by using genomic DNA of Toxoplasma gondii as template; and ( B ) colony PCR confirmation for cloning ROP8 in <t>pVAX-1-GFP</t> vector; ( C ) colony PCR confirmation for
    Pvax 1 Vector, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 131 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pvax 1 vector/product/Thermo Fisher
    Average 99 stars, based on 131 article reviews
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    93
    TaKaRa pvax1 vector
    Salivary PAc-specific IgA and serum PAc-specific IgG antibody levels in rats immunized intranasally with pCIA-P in combination with pCCL17/VAX, pCCL19/VAX, pCCL17/VAX plus pCCL19/VAX, or <t>pVAX1.</t> Saliva and serum samples were collected 52 days after the first immunization. The specific anti-PAc salivary IgA (A) and specific anti-PAc serum IgG (B) concentrations were determined by ELISA. The data are expressed as the mean±SD. n =5. * P
    Pvax1 Vector, supplied by TaKaRa, used in various techniques. Bioz Stars score: 93/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    GenScript pvax1 expression vector
    Salivary PAc-specific IgA and serum PAc-specific IgG antibody levels in rats immunized intranasally with pCIA-P in combination with pCCL17/VAX, pCCL19/VAX, pCCL17/VAX plus pCCL19/VAX, or <t>pVAX1.</t> Saliva and serum samples were collected 52 days after the first immunization. The specific anti-PAc salivary IgA (A) and specific anti-PAc serum IgG (B) concentrations were determined by ELISA. The data are expressed as the mean±SD. n =5. * P
    Pvax1 Expression Vector, supplied by GenScript, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pvax1 expression vector/product/GenScript
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pvax1 expression vector - by Bioz Stars, 2020-04
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    93
    GenScript pvax1 vector
    Salivary PAc-specific IgA and serum PAc-specific IgG antibody levels in rats immunized intranasally with pCIA-P in combination with pCCL17/VAX, pCCL19/VAX, pCCL17/VAX plus pCCL19/VAX, or <t>pVAX1.</t> Saliva and serum samples were collected 52 days after the first immunization. The specific anti-PAc salivary IgA (A) and specific anti-PAc serum IgG (B) concentrations were determined by ELISA. The data are expressed as the mean±SD. n =5. * P
    Pvax1 Vector, supplied by GenScript, used in various techniques. Bioz Stars score: 93/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pvax1 vector/product/GenScript
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    91
    Millipore pvax1 expression vector
    Restriction Enzyme Analysis of Recombinant <t>pVAX1-SAG1</t> Line 1 and 2, the recombinant pVAX-SAG1 plasmid digested by NheI; Line 3, 1 kb DNA ladder; Line 4, the pVAX vector digested by NheI.
    Pvax1 Expression Vector, supplied by Millipore, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    87
    Thermo Fisher ecori digested pvax1 mammalian expression vector
    Restriction Enzyme Analysis of Recombinant <t>pVAX1-SAG1</t> Line 1 and 2, the recombinant pVAX-SAG1 plasmid digested by NheI; Line 3, 1 kb DNA ladder; Line 4, the pVAX vector digested by NheI.
    Ecori Digested Pvax1 Mammalian Expression Vector, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 87/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ecori digested pvax1 mammalian expression vector/product/Thermo Fisher
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    94
    Vical pvax1 mammalian expression vector
    Restriction Enzyme Analysis of Recombinant <t>pVAX1-SAG1</t> Line 1 and 2, the recombinant pVAX-SAG1 plasmid digested by NheI; Line 3, 1 kb DNA ladder; Line 4, the pVAX vector digested by NheI.
    Pvax1 Mammalian Expression Vector, supplied by Vical, used in various techniques. Bioz Stars score: 94/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Thermo Fisher mammalian expression vectors pvax1
    Restriction Enzyme Analysis of Recombinant <t>pVAX1-SAG1</t> Line 1 and 2, the recombinant pVAX-SAG1 plasmid digested by NheI; Line 3, 1 kb DNA ladder; Line 4, the pVAX vector digested by NheI.
    Mammalian Expression Vectors Pvax1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Thermo Fisher zfn expression plasmid pvax1
    Restriction Enzyme Analysis of Recombinant <t>pVAX1-SAG1</t> Line 1 and 2, the recombinant pVAX-SAG1 plasmid digested by NheI; Line 3, 1 kb DNA ladder; Line 4, the pVAX vector digested by NheI.
    Zfn Expression Plasmid Pvax1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Thermo Fisher prmt6 expression plasmid pvax1
    Restriction Enzyme Analysis of Recombinant <t>pVAX1-SAG1</t> Line 1 and 2, the recombinant pVAX-SAG1 plasmid digested by NheI; Line 3, 1 kb DNA ladder; Line 4, the pVAX vector digested by NheI.
    Prmt6 Expression Plasmid Pvax1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Agarose gel electrophoresis for ( A ) polymerase chain reaction (PCR) amplification of ROP8 gene by using genomic DNA of Toxoplasma gondii as template; and ( B ) colony PCR confirmation for cloning ROP8 in pVAX-1-GFP vector; ( C ) colony PCR confirmation for

    Journal: The American Journal of Tropical Medicine and Hygiene

    Article Title: Protective Immune Response in BALB/c Mice Induced by DNA Vaccine of the ROP8 gene of Toxoplasma gondii

    doi: 10.4269/ajtmh.12-0727

    Figure Lengend Snippet: Agarose gel electrophoresis for ( A ) polymerase chain reaction (PCR) amplification of ROP8 gene by using genomic DNA of Toxoplasma gondii as template; and ( B ) colony PCR confirmation for cloning ROP8 in pVAX-1-GFP vector; ( C ) colony PCR confirmation for

    Article Snippet: The ROP8 gene was digested with Hind III and EcoR 1 and ligated into the pVAX-1 vector (Invitrogen, USA) digested with the same restriction enzymes.

    Techniques: Agarose Gel Electrophoresis, Polymerase Chain Reaction, Amplification, Clone Assay, Plasmid Preparation

    Detection of specific anti-ROP8 antibodies in sera of mice immunized with ROP8 -pVAX-1, by western blotting using Toxoplasma gondii antigen. Reactivity of sera from mice immunized with, ROP8 -pVAX-1 (lanes 1 and 2), empty plasmid pVAX-1 (lanes 3 and 4),

    Journal: The American Journal of Tropical Medicine and Hygiene

    Article Title: Protective Immune Response in BALB/c Mice Induced by DNA Vaccine of the ROP8 gene of Toxoplasma gondii

    doi: 10.4269/ajtmh.12-0727

    Figure Lengend Snippet: Detection of specific anti-ROP8 antibodies in sera of mice immunized with ROP8 -pVAX-1, by western blotting using Toxoplasma gondii antigen. Reactivity of sera from mice immunized with, ROP8 -pVAX-1 (lanes 1 and 2), empty plasmid pVAX-1 (lanes 3 and 4),

    Article Snippet: The ROP8 gene was digested with Hind III and EcoR 1 and ligated into the pVAX-1 vector (Invitrogen, USA) digested with the same restriction enzymes.

    Techniques: Mouse Assay, Western Blot, Plasmid Preparation

    The in vitro proliferation of splenocytes from mice vaccinated with ROP8 -pVAX-1 after stimulation with total lysate antigen (TLA). Splenocytes from mice immunized with pVAX-1, PBS, ROP8 -pVAX-1 were harvested 2 weeks after the third DNA booster. Following

    Journal: The American Journal of Tropical Medicine and Hygiene

    Article Title: Protective Immune Response in BALB/c Mice Induced by DNA Vaccine of the ROP8 gene of Toxoplasma gondii

    doi: 10.4269/ajtmh.12-0727

    Figure Lengend Snippet: The in vitro proliferation of splenocytes from mice vaccinated with ROP8 -pVAX-1 after stimulation with total lysate antigen (TLA). Splenocytes from mice immunized with pVAX-1, PBS, ROP8 -pVAX-1 were harvested 2 weeks after the third DNA booster. Following

    Article Snippet: The ROP8 gene was digested with Hind III and EcoR 1 and ligated into the pVAX-1 vector (Invitrogen, USA) digested with the same restriction enzymes.

    Techniques: In Vitro, Mouse Assay

    Immunoblot analysis of expression and antigenicity of ROP8 protein. Lysate of Chinese hamster ovary (CHO) cells transfected with pVAX-1-GFP (lanes 1 and 2), ROP8 -pVAX-1-GFP (lanes 3 and 4), mock transfection (lanes 5 and 6), ROP8 -pVAX-1 (lanes 7 and 8)

    Journal: The American Journal of Tropical Medicine and Hygiene

    Article Title: Protective Immune Response in BALB/c Mice Induced by DNA Vaccine of the ROP8 gene of Toxoplasma gondii

    doi: 10.4269/ajtmh.12-0727

    Figure Lengend Snippet: Immunoblot analysis of expression and antigenicity of ROP8 protein. Lysate of Chinese hamster ovary (CHO) cells transfected with pVAX-1-GFP (lanes 1 and 2), ROP8 -pVAX-1-GFP (lanes 3 and 4), mock transfection (lanes 5 and 6), ROP8 -pVAX-1 (lanes 7 and 8)

    Article Snippet: The ROP8 gene was digested with Hind III and EcoR 1 and ligated into the pVAX-1 vector (Invitrogen, USA) digested with the same restriction enzymes.

    Techniques: Expressing, Transfection

    Expression analyses of ROP8 protein in transfected Chinese hamster ovary (CHO) cells at 72 h post-transfection ( A ) ROP8 -pVAX-1-GFP protein expressed in CHO cells; ( B ) expression of pVAX-1-GFP as positive control; ( C ) transfection with no DNA as negative

    Journal: The American Journal of Tropical Medicine and Hygiene

    Article Title: Protective Immune Response in BALB/c Mice Induced by DNA Vaccine of the ROP8 gene of Toxoplasma gondii

    doi: 10.4269/ajtmh.12-0727

    Figure Lengend Snippet: Expression analyses of ROP8 protein in transfected Chinese hamster ovary (CHO) cells at 72 h post-transfection ( A ) ROP8 -pVAX-1-GFP protein expressed in CHO cells; ( B ) expression of pVAX-1-GFP as positive control; ( C ) transfection with no DNA as negative

    Article Snippet: The ROP8 gene was digested with Hind III and EcoR 1 and ligated into the pVAX-1 vector (Invitrogen, USA) digested with the same restriction enzymes.

    Techniques: Expressing, Transfection, Positive Control

    Survival rate of mice immunized with ROP8 -pVAX-1 (▲), pVAX-1 (●), and phosphate buffered saline (PBS) (■) when challenged with 1 × 10 3 tachyzoites, the number of mice for each group is 12.

    Journal: The American Journal of Tropical Medicine and Hygiene

    Article Title: Protective Immune Response in BALB/c Mice Induced by DNA Vaccine of the ROP8 gene of Toxoplasma gondii

    doi: 10.4269/ajtmh.12-0727

    Figure Lengend Snippet: Survival rate of mice immunized with ROP8 -pVAX-1 (▲), pVAX-1 (●), and phosphate buffered saline (PBS) (■) when challenged with 1 × 10 3 tachyzoites, the number of mice for each group is 12.

    Article Snippet: The ROP8 gene was digested with Hind III and EcoR 1 and ligated into the pVAX-1 vector (Invitrogen, USA) digested with the same restriction enzymes.

    Techniques: Mouse Assay

    Production of IFN-γ and IL-4 by splenocytes from mice vaccinated with ROP8 -pVAX-1. Data represent the mean production of IFN-γ ( A ) and IL-4 ( B ) as determined by enzyme-linked immunosorbent assay (ELISA) of mice vaccinated with phosphate

    Journal: The American Journal of Tropical Medicine and Hygiene

    Article Title: Protective Immune Response in BALB/c Mice Induced by DNA Vaccine of the ROP8 gene of Toxoplasma gondii

    doi: 10.4269/ajtmh.12-0727

    Figure Lengend Snippet: Production of IFN-γ and IL-4 by splenocytes from mice vaccinated with ROP8 -pVAX-1. Data represent the mean production of IFN-γ ( A ) and IL-4 ( B ) as determined by enzyme-linked immunosorbent assay (ELISA) of mice vaccinated with phosphate

    Article Snippet: The ROP8 gene was digested with Hind III and EcoR 1 and ligated into the pVAX-1 vector (Invitrogen, USA) digested with the same restriction enzymes.

    Techniques: Mouse Assay, Enzyme-linked Immunosorbent Assay

    Protection against K. pneumoniae afforded upon active immunization of mice. The survival of challenged mice vaccinated with either of the DNA vaccines by the intradermal (A) or intramuscular (B) route was compared. Mice were challenged intraperitoneally with 10 8 CFU (the LD) of K. pneumoniae. At day 8, mice given the pOmpA ( P = 0.0081) or pOmpK36 ( P = 0.0032) DNA vaccine by the intradermal route had a significantly higher degree of protection than mice from the control groups receiving PBS or the pVAX1 plasmid DNA. P values were calculated by Kaplan-Meier survival curves and the log rank test.

    Journal: Clinical and Vaccine Immunology : CVI

    Article Title: Protective Efficacy of DNA Vaccines Encoding Outer Membrane Protein A and OmpK36 of Klebsiella pneumoniae in Mice ▿

    doi: 10.1128/CVI.00275-10

    Figure Lengend Snippet: Protection against K. pneumoniae afforded upon active immunization of mice. The survival of challenged mice vaccinated with either of the DNA vaccines by the intradermal (A) or intramuscular (B) route was compared. Mice were challenged intraperitoneally with 10 8 CFU (the LD) of K. pneumoniae. At day 8, mice given the pOmpA ( P = 0.0081) or pOmpK36 ( P = 0.0032) DNA vaccine by the intradermal route had a significantly higher degree of protection than mice from the control groups receiving PBS or the pVAX1 plasmid DNA. P values were calculated by Kaplan-Meier survival curves and the log rank test.

    Article Snippet: The restriction endonuclease (RE)-digested PCR products and the pVAX1 vector were ligated with T4 DNA ligase (Life Technologies, Gaithersburg, MD).

    Techniques: Mouse Assay, Plasmid Preparation

    In vivo kinetics of reporter signal after co-delivery of luciferase, and HIV PR and RT genes. Delivery and expression of HIV-1 protease (PR) and reverse transcriptase (RT) encoded by their expression-optimized genes in pVax1 vector (PR, RT) monitored indirectly by their co-administration with a plasmid directing the expression of firefly luciferase pVaxLuc (Luc). Mice (n = 5 per group) were immunized by ID or IM injections of PR/Luc, or RT/Luc, or pVax1/Luc, 10 μ ] using a DermaVax electroporator equipped with multi-needle electrodes (Cellectis, Paris, France). Total photon flux from the injection sites was assessed by BLI on days 1, 3, 9, 15 and 21 as described in the Materials and methods. Data represent individual values for each injection site, and mean values (A). Luminescence kinetics measured by 2D BLI after delivery of PR and RT by ID (B) and IM routes (C). Luminescence kinetics was also registered by 3D BLI demonstrating the volume of expressing tissues after PR/Luc (D), RT/Luc (E) and vector/Luc administration (F). Data represent average photon flux (photons/sq cm/sec) and expression volume (mm 3 ) for 4 to 5 mice per group and time point, with two simultaneous measurements per mouse. Statistical comparison was done using Mann Whitney U-test; * p

    Journal: PLoS ONE

    Article Title: DNA immunization site determines the level of gene expression and the magnitude, but not the type of the induced immune response

    doi: 10.1371/journal.pone.0197902

    Figure Lengend Snippet: In vivo kinetics of reporter signal after co-delivery of luciferase, and HIV PR and RT genes. Delivery and expression of HIV-1 protease (PR) and reverse transcriptase (RT) encoded by their expression-optimized genes in pVax1 vector (PR, RT) monitored indirectly by their co-administration with a plasmid directing the expression of firefly luciferase pVaxLuc (Luc). Mice (n = 5 per group) were immunized by ID or IM injections of PR/Luc, or RT/Luc, or pVax1/Luc, 10 μ ] using a DermaVax electroporator equipped with multi-needle electrodes (Cellectis, Paris, France). Total photon flux from the injection sites was assessed by BLI on days 1, 3, 9, 15 and 21 as described in the Materials and methods. Data represent individual values for each injection site, and mean values (A). Luminescence kinetics measured by 2D BLI after delivery of PR and RT by ID (B) and IM routes (C). Luminescence kinetics was also registered by 3D BLI demonstrating the volume of expressing tissues after PR/Luc (D), RT/Luc (E) and vector/Luc administration (F). Data represent average photon flux (photons/sq cm/sec) and expression volume (mm 3 ) for 4 to 5 mice per group and time point, with two simultaneous measurements per mouse. Statistical comparison was done using Mann Whitney U-test; * p

    Article Snippet: The luciferase-coding plasmid, pVax-luc 4663 bp (pVaxLuc) constructed by inserting the cDNA of firefly luciferase from pGL2-basic vector (Promega, # E1641) into vector pVAX1 (Invitrogen, # V260-20) under the control of a human cytomegalovirus immediate/early promoter and a polyadenylation signal from the bovine growth hormone gene [ ], was kindly provided by Maltais AK (Karolinska Institutet, currently Eurocine Vaccines AB, Sweden).

    Techniques: In Vivo, Luciferase, Expressing, Plasmid Preparation, Mouse Assay, Injection, Size-exclusion Chromatography, MANN-WHITNEY

    In vitro transcription activity of man-PEI and PEI 25k on DC 2.4 cells. ( A ) Quantified by β-galactosidase assay using plasmid encoding lacZ as a reporter gene. ( B ) Transfected with pVAX1-HIV gag and quantified by the transcription level of HIV gag gene using real-time PCR. Notes: The β-galactosidase activity of the m-PEI/DNA group was significantly higher than that of the naked DNA group and the PEI 25k/DNA group, *** P

    Journal: International Journal of Nanomedicine

    Article Title: Enhanced immune response against HIV-1 induced by a heterologous DNA prime-adenovirus boost vaccination using mannosylated polyethyleneimine as DNA vaccine adjuvant

    doi: 10.2147/IJN.S43827

    Figure Lengend Snippet: In vitro transcription activity of man-PEI and PEI 25k on DC 2.4 cells. ( A ) Quantified by β-galactosidase assay using plasmid encoding lacZ as a reporter gene. ( B ) Transfected with pVAX1-HIV gag and quantified by the transcription level of HIV gag gene using real-time PCR. Notes: The β-galactosidase activity of the m-PEI/DNA group was significantly higher than that of the naked DNA group and the PEI 25k/DNA group, *** P

    Article Snippet: The PCR product was then digested by restriction enzymes Xba 1 and Hind 3, and inserted into backbone vector pVAX1 (Life Technologies, Carlsbad, CA, USA), which was digested by the same enzymes.

    Techniques: In Vitro, Activity Assay, Plasmid Preparation, Transfection, Real-time Polymerase Chain Reaction

    Agarose gel electrophoresis of pVAX1-HIV gag and pVAX1 vectors. Notes: The empty pVAX1 vector was 3000 bp. After insertion of HIV gag fragment (1500 bp), the pVAX1-HIV gag was 4500 bp. Abbreviation: bp, base pair; HIV, human immunodeficiency virus.

    Journal: International Journal of Nanomedicine

    Article Title: Enhanced immune response against HIV-1 induced by a heterologous DNA prime-adenovirus boost vaccination using mannosylated polyethyleneimine as DNA vaccine adjuvant

    doi: 10.2147/IJN.S43827

    Figure Lengend Snippet: Agarose gel electrophoresis of pVAX1-HIV gag and pVAX1 vectors. Notes: The empty pVAX1 vector was 3000 bp. After insertion of HIV gag fragment (1500 bp), the pVAX1-HIV gag was 4500 bp. Abbreviation: bp, base pair; HIV, human immunodeficiency virus.

    Article Snippet: The PCR product was then digested by restriction enzymes Xba 1 and Hind 3, and inserted into backbone vector pVAX1 (Life Technologies, Carlsbad, CA, USA), which was digested by the same enzymes.

    Techniques: Agarose Gel Electrophoresis, Plasmid Preparation

    Construction and characterization of pIA. ( A ) These DNA sequences were subcloned into the Kpn I, BamH I and Xho I sites of pVAX1 to construct pVAX1-IL22 (pIL-22) and pVAX1-IA (pIA) respectively. ( B ) Expression of the IL-22 and ApoA-I fusion protein was detected by western blot from the indicated amount in HEK-293T cells after transfection with pIA, pVAX1 vector, or PBS (control group). ( C ) Densitometric values were quantified and normalized to control ( n = 3; mean ± SD; ** P

    Journal: Theranostics

    Article Title: Tethering Interleukin-22 to Apolipoprotein A-I Ameliorates Mice from Acetaminophen-induced Liver Injury

    doi: 10.7150/thno.20955

    Figure Lengend Snippet: Construction and characterization of pIA. ( A ) These DNA sequences were subcloned into the Kpn I, BamH I and Xho I sites of pVAX1 to construct pVAX1-IL22 (pIL-22) and pVAX1-IA (pIA) respectively. ( B ) Expression of the IL-22 and ApoA-I fusion protein was detected by western blot from the indicated amount in HEK-293T cells after transfection with pIA, pVAX1 vector, or PBS (control group). ( C ) Densitometric values were quantified and normalized to control ( n = 3; mean ± SD; ** P

    Article Snippet: Experiments were performed using pVAX1 vectors (Invitrogen, USA).

    Techniques: Construct, IA, Expressing, Western Blot, Transfection, Plasmid Preparation

    Interferon-γ response to CHIKV envelope E2 measured by ELISpot. C57BL/6 mice were immunized two times, each 2 weeks apart, with 25 µg pVax1 vector or pCHIKV-E2 and sacrificed 1 week later. (A) Splenocytes were harvested and cultured overnight

    Journal:

    Article Title: IMMUNOGENICITY OF NOVEL CONSENSUS-BASED DNA VACCINES AGAINST CHIKUNGUNYA VIRUS

    doi: 10.1016/j.vaccine.2008.03.060

    Figure Lengend Snippet: Interferon-γ response to CHIKV envelope E2 measured by ELISpot. C57BL/6 mice were immunized two times, each 2 weeks apart, with 25 µg pVax1 vector or pCHIKV-E2 and sacrificed 1 week later. (A) Splenocytes were harvested and cultured overnight

    Article Snippet: Consensus sequences were optimized for expression, including codon and RNA optimization (GeneArt, Regensburg, Germany) and inserted into the pVax1 expression vector (Invitrogen).

    Techniques: Enzyme-linked Immunospot, Mouse Assay, Plasmid Preparation, Cell Culture

    Anitbody ELISA. (A) C57BL/6 mice were immunized two times, each 2 weeks apart, with 25µg 25µg pVax1 vector or CHIKV plasmids as indicated and sacrificed 1 week later. Serum was collected and subject to analysis for Total IgG production.

    Journal:

    Article Title: IMMUNOGENICITY OF NOVEL CONSENSUS-BASED DNA VACCINES AGAINST CHIKUNGUNYA VIRUS

    doi: 10.1016/j.vaccine.2008.03.060

    Figure Lengend Snippet: Anitbody ELISA. (A) C57BL/6 mice were immunized two times, each 2 weeks apart, with 25µg 25µg pVax1 vector or CHIKV plasmids as indicated and sacrificed 1 week later. Serum was collected and subject to analysis for Total IgG production.

    Article Snippet: Consensus sequences were optimized for expression, including codon and RNA optimization (GeneArt, Regensburg, Germany) and inserted into the pVax1 expression vector (Invitrogen).

    Techniques: Enzyme-linked Immunosorbent Assay, Mouse Assay, Plasmid Preparation

    Interferon-γ response to envelope E1 measured by ELISpot. C57BL/6 mice were immunized two times, each 2 weeks apart, with 25µg pVax1 vector or pCHIKV-E1 and sacrificed 1 week later. (A) Splenocytes were harvested and cultured overnight

    Journal:

    Article Title: IMMUNOGENICITY OF NOVEL CONSENSUS-BASED DNA VACCINES AGAINST CHIKUNGUNYA VIRUS

    doi: 10.1016/j.vaccine.2008.03.060

    Figure Lengend Snippet: Interferon-γ response to envelope E1 measured by ELISpot. C57BL/6 mice were immunized two times, each 2 weeks apart, with 25µg pVax1 vector or pCHIKV-E1 and sacrificed 1 week later. (A) Splenocytes were harvested and cultured overnight

    Article Snippet: Consensus sequences were optimized for expression, including codon and RNA optimization (GeneArt, Regensburg, Germany) and inserted into the pVax1 expression vector (Invitrogen).

    Techniques: Enzyme-linked Immunospot, Mouse Assay, Plasmid Preparation, Cell Culture

    Interferon-γ response CHIKV-Capsid measured by ELISpot. C57BL/6 mice were immunized two times, each 2 weeks apart, with 25µg pVax1 vector or pCHIKV-Capsid and sacrificed 1 week later. (A) Splenocytes were harvested and cultured overnight

    Journal:

    Article Title: IMMUNOGENICITY OF NOVEL CONSENSUS-BASED DNA VACCINES AGAINST CHIKUNGUNYA VIRUS

    doi: 10.1016/j.vaccine.2008.03.060

    Figure Lengend Snippet: Interferon-γ response CHIKV-Capsid measured by ELISpot. C57BL/6 mice were immunized two times, each 2 weeks apart, with 25µg pVax1 vector or pCHIKV-Capsid and sacrificed 1 week later. (A) Splenocytes were harvested and cultured overnight

    Article Snippet: Consensus sequences were optimized for expression, including codon and RNA optimization (GeneArt, Regensburg, Germany) and inserted into the pVax1 expression vector (Invitrogen).

    Techniques: Enzyme-linked Immunospot, Mouse Assay, Plasmid Preparation, Cell Culture

    (A) Schematic representation of the strategy for cloning the IgE–leader CHIKV fusion gene into the pVax1 vector. (B) Agarose gel photograph showing the CHIKV plasmid (Envelope E1, E2 and Capsid) linear specific band indicated (lane 4) with Kpn1

    Journal:

    Article Title: IMMUNOGENICITY OF NOVEL CONSENSUS-BASED DNA VACCINES AGAINST CHIKUNGUNYA VIRUS

    doi: 10.1016/j.vaccine.2008.03.060

    Figure Lengend Snippet: (A) Schematic representation of the strategy for cloning the IgE–leader CHIKV fusion gene into the pVax1 vector. (B) Agarose gel photograph showing the CHIKV plasmid (Envelope E1, E2 and Capsid) linear specific band indicated (lane 4) with Kpn1

    Article Snippet: Consensus sequences were optimized for expression, including codon and RNA optimization (GeneArt, Regensburg, Germany) and inserted into the pVax1 expression vector (Invitrogen).

    Techniques: Clone Assay, Plasmid Preparation, Agarose Gel Electrophoresis

    Identification of the multivalent DNA plasmid pPCFN-CpG. a Schematic representation of pPCFN-CpG. The chimeric gene containing four different virulence genes was inserted into pVAX1 utilizing Hin dIII and Eco RV sites. b The multivalent DNA plasmids were separated by electrophoresis. Lane 1, pVAX1 empty plasmid; Lane 2, pPCFN-CpG plasmid; Lane M, DNA marker-DL5000. c The digestion products were separated by electrophoresis. Lane 1, two fragments after restriction enzyme digestion with Hin dIII and Eco RV; Lane 2, pPCFN-CpG plasmid. Lane M, DNA marker-DL5000. The position of the 1482 bp band is indicated by the arrow. pPCFN-CpG, the DNA plasmid contains the epitope of plo , cbpA , fimA , and nanH gene of T. pyogenes and CpG ODN1826 motif

    Journal: Journal of Nanobiotechnology

    Article Title: Chitosan-DNA nanoparticles enhanced the immunogenicity of multivalent DNA vaccination on mice against Trueperella pyogenes infection

    doi: 10.1186/s12951-018-0337-2

    Figure Lengend Snippet: Identification of the multivalent DNA plasmid pPCFN-CpG. a Schematic representation of pPCFN-CpG. The chimeric gene containing four different virulence genes was inserted into pVAX1 utilizing Hin dIII and Eco RV sites. b The multivalent DNA plasmids were separated by electrophoresis. Lane 1, pVAX1 empty plasmid; Lane 2, pPCFN-CpG plasmid; Lane M, DNA marker-DL5000. c The digestion products were separated by electrophoresis. Lane 1, two fragments after restriction enzyme digestion with Hin dIII and Eco RV; Lane 2, pPCFN-CpG plasmid. Lane M, DNA marker-DL5000. The position of the 1482 bp band is indicated by the arrow. pPCFN-CpG, the DNA plasmid contains the epitope of plo , cbpA , fimA , and nanH gene of T. pyogenes and CpG ODN1826 motif

    Article Snippet: The multi-epitope chimeric gene was then incorporated into expression vector pVAX1 (Invitrogen, USA) designated as pPCFN-CpG, which was confirmed by endonuclease digestion assay and DNA sequencing.

    Techniques: Plasmid Preparation, Electrophoresis, Marker

    Characterization and stability of the pPCFN-CpG-CS-NPs. Transmission electron microscopy micrograph of the pPFCN-CpG-CS-NPs (magnification 30,000×). a pPCFN-CpG-CS-NPs at pH 5.5 is indicated by the arrow. b – d Measurement of these particles showed a narrow distribution of the pPCFN-CpG-CS-NPs, and the average diameter was 93.58 nm. e Measurement of these particles showed a Zeta potential of + 5.27 mV. f Stability analysis of the plasmid DNA after encapsulation in the chitosan nanoparticles. Lane 1: pPCFN-CpG-CS-NPs treated by DNase I and chitosanase; Lane 2: pPCFN-CpG-CS-NPs treated by DNase I; Lane 3: untreated chitosan encapsulated plasmid pPCFN-CpG; Lane4: untreated naked plasmid pPCFN-CpG; Lane 5: pPCFN-CS-NPs treated by DNase I; Lane 6: naked plasmid pVAX1-PCFN treated by DNase I; M: DNA marker DL 5000. pPCFN-CpG-CS-NPs, the pPCFN-CpG DNA plasmid encapsulated in chitosan nanoparticles; pPCFN-CS-NPs, the pVAX1-PCFN DNA plasmid encapsulated in chitosan nanoparticles

    Journal: Journal of Nanobiotechnology

    Article Title: Chitosan-DNA nanoparticles enhanced the immunogenicity of multivalent DNA vaccination on mice against Trueperella pyogenes infection

    doi: 10.1186/s12951-018-0337-2

    Figure Lengend Snippet: Characterization and stability of the pPCFN-CpG-CS-NPs. Transmission electron microscopy micrograph of the pPFCN-CpG-CS-NPs (magnification 30,000×). a pPCFN-CpG-CS-NPs at pH 5.5 is indicated by the arrow. b – d Measurement of these particles showed a narrow distribution of the pPCFN-CpG-CS-NPs, and the average diameter was 93.58 nm. e Measurement of these particles showed a Zeta potential of + 5.27 mV. f Stability analysis of the plasmid DNA after encapsulation in the chitosan nanoparticles. Lane 1: pPCFN-CpG-CS-NPs treated by DNase I and chitosanase; Lane 2: pPCFN-CpG-CS-NPs treated by DNase I; Lane 3: untreated chitosan encapsulated plasmid pPCFN-CpG; Lane4: untreated naked plasmid pPCFN-CpG; Lane 5: pPCFN-CS-NPs treated by DNase I; Lane 6: naked plasmid pVAX1-PCFN treated by DNase I; M: DNA marker DL 5000. pPCFN-CpG-CS-NPs, the pPCFN-CpG DNA plasmid encapsulated in chitosan nanoparticles; pPCFN-CS-NPs, the pVAX1-PCFN DNA plasmid encapsulated in chitosan nanoparticles

    Article Snippet: The multi-epitope chimeric gene was then incorporated into expression vector pVAX1 (Invitrogen, USA) designated as pPCFN-CpG, which was confirmed by endonuclease digestion assay and DNA sequencing.

    Techniques: Transmission Assay, Electron Microscopy, Plasmid Preparation, Marker

    Humoral immune responses induced by the plasmids. Mice were immunized with plasmid pPCFN-CpG, pVAX1-PCFN, pVAX1-PCF, pVAX1-PC, or pPCFN-CpG-CS-NPs respectively, and boosted once with a 3-week interval. PBS was used as a control. Serum samples were collected from the tail vein of mice at the indicated time points. The titers of the PLO-specific antibody were detected by indirect ELISA assay. a Serum total IgG titers against PLO in immunized mice; b serum IgG1 titers against PLO in immunized mice; c serum IgG2a titers against PLO in immunized mice; d the dynamic changes of serum IgG2a/IgG1 against PLO in immunized mice. Data represented mean ± SD of OD at 490 nm. * p

    Journal: Journal of Nanobiotechnology

    Article Title: Chitosan-DNA nanoparticles enhanced the immunogenicity of multivalent DNA vaccination on mice against Trueperella pyogenes infection

    doi: 10.1186/s12951-018-0337-2

    Figure Lengend Snippet: Humoral immune responses induced by the plasmids. Mice were immunized with plasmid pPCFN-CpG, pVAX1-PCFN, pVAX1-PCF, pVAX1-PC, or pPCFN-CpG-CS-NPs respectively, and boosted once with a 3-week interval. PBS was used as a control. Serum samples were collected from the tail vein of mice at the indicated time points. The titers of the PLO-specific antibody were detected by indirect ELISA assay. a Serum total IgG titers against PLO in immunized mice; b serum IgG1 titers against PLO in immunized mice; c serum IgG2a titers against PLO in immunized mice; d the dynamic changes of serum IgG2a/IgG1 against PLO in immunized mice. Data represented mean ± SD of OD at 490 nm. * p

    Article Snippet: The multi-epitope chimeric gene was then incorporated into expression vector pVAX1 (Invitrogen, USA) designated as pPCFN-CpG, which was confirmed by endonuclease digestion assay and DNA sequencing.

    Techniques: Mouse Assay, Plasmid Preparation, Indirect ELISA

    Transient expression of the chimeric protein and activation of CpG. a HEK293T cells were transfected with plasmid pPCFN-CpG, pVAX1-PCFN, pVAX1-PCF, pVAX1-PC, pVAX1, or pPCFN-CpG-CS-NPs respectively. Transient expression of proteins was detected with anti-PLO mouse polyclonal antibody by using fluorescence microscopy assays (A3-A7). (A1) Cell controls. (A2) Cells were transfected with vector pVAX1. FITC: FITC-conjugated goat anti-mouse IgG; Hoechst: cell nuclei; Merge: the overlay fluorescence images of FITC and nucleus; Bright: the external profiles of HEK293T cells. The scale bar is 50 μm. b and c RAW264.7 were transfected with pVAX1, pVAX1-PCFN or pPCFN-CpG plasmid for 36 h. The expression levels of TLR9 and Myd88 were detected by western blotting ( b ) and qPCR ( c ). Data are shown as the mean ± SEM of three independent experiments. *** p

    Journal: Journal of Nanobiotechnology

    Article Title: Chitosan-DNA nanoparticles enhanced the immunogenicity of multivalent DNA vaccination on mice against Trueperella pyogenes infection

    doi: 10.1186/s12951-018-0337-2

    Figure Lengend Snippet: Transient expression of the chimeric protein and activation of CpG. a HEK293T cells were transfected with plasmid pPCFN-CpG, pVAX1-PCFN, pVAX1-PCF, pVAX1-PC, pVAX1, or pPCFN-CpG-CS-NPs respectively. Transient expression of proteins was detected with anti-PLO mouse polyclonal antibody by using fluorescence microscopy assays (A3-A7). (A1) Cell controls. (A2) Cells were transfected with vector pVAX1. FITC: FITC-conjugated goat anti-mouse IgG; Hoechst: cell nuclei; Merge: the overlay fluorescence images of FITC and nucleus; Bright: the external profiles of HEK293T cells. The scale bar is 50 μm. b and c RAW264.7 were transfected with pVAX1, pVAX1-PCFN or pPCFN-CpG plasmid for 36 h. The expression levels of TLR9 and Myd88 were detected by western blotting ( b ) and qPCR ( c ). Data are shown as the mean ± SEM of three independent experiments. *** p

    Article Snippet: The multi-epitope chimeric gene was then incorporated into expression vector pVAX1 (Invitrogen, USA) designated as pPCFN-CpG, which was confirmed by endonuclease digestion assay and DNA sequencing.

    Techniques: Expressing, Activation Assay, Transfection, Plasmid Preparation, Fluorescence, Microscopy, Western Blot, Real-time Polymerase Chain Reaction

    Effect of pVAX1-MUC1-VNTR n DNA vaccine immunization on panc02-MUC1 tumor growth in mice. The pVAX1-MUC1-VNTR 6 DNA vaccine showed a stronger inhibitory effect on panc02-MUC1 tumor growth in vivo compared with the strong pVAX1-MUC1-VNTR1, VNTR 3 , VNTR 4 and VNTR 9 groups. pVAX1-MUC1-VNTR n DNA vaccines showed no evident inhibitory effect on panc02 tumor cells, indicating MUC1 specificity. *P

    Journal: Oncology Letters

    Article Title: Optimized construction of MUC1-VNTRn DNA vaccine and its anti-pancreatic cancer efficacy

    doi: 10.3892/ol.2017.5717

    Figure Lengend Snippet: Effect of pVAX1-MUC1-VNTR n DNA vaccine immunization on panc02-MUC1 tumor growth in mice. The pVAX1-MUC1-VNTR 6 DNA vaccine showed a stronger inhibitory effect on panc02-MUC1 tumor growth in vivo compared with the strong pVAX1-MUC1-VNTR1, VNTR 3 , VNTR 4 and VNTR 9 groups. pVAX1-MUC1-VNTR n DNA vaccines showed no evident inhibitory effect on panc02 tumor cells, indicating MUC1 specificity. *P

    Article Snippet: A total of three groups were classified: MUC1-VNTRn group; pVAX1 without vector group; and HeLa cells group (negative control group).

    Techniques: Mouse Assay, In Vivo

    Effect of treatment with thepVAX1-MUC1-VNTR n DNA vaccines on in vivo growth of panc02-MUC1 tumors. The pVAX1-MUC1-VNTR 6 DNA vaccine showed a stronger inhibitory effect on panc02-MUC1 tumor growth compared with the strong pVAX1-MUC1-VNTR 1 , VNTR 3 , VNTR 4 and VNTR 9 groups. No evident inhibitory effects on panc02 tumor cells were observed for all pVAX1-MUC1-VNTR n DNA vaccines, indicating MUC1 specificity. *P

    Journal: Oncology Letters

    Article Title: Optimized construction of MUC1-VNTRn DNA vaccine and its anti-pancreatic cancer efficacy

    doi: 10.3892/ol.2017.5717

    Figure Lengend Snippet: Effect of treatment with thepVAX1-MUC1-VNTR n DNA vaccines on in vivo growth of panc02-MUC1 tumors. The pVAX1-MUC1-VNTR 6 DNA vaccine showed a stronger inhibitory effect on panc02-MUC1 tumor growth compared with the strong pVAX1-MUC1-VNTR 1 , VNTR 3 , VNTR 4 and VNTR 9 groups. No evident inhibitory effects on panc02 tumor cells were observed for all pVAX1-MUC1-VNTR n DNA vaccines, indicating MUC1 specificity. *P

    Article Snippet: A total of three groups were classified: MUC1-VNTRn group; pVAX1 without vector group; and HeLa cells group (negative control group).

    Techniques: In Vivo

    Inhibition of BRAF V600E in induced tumors led to tumor outgrowth inhibition. Melanomas were induced by 4-hydroxytamoxifen application on the flank of 4–10-week-old C57BL/6J Tyr::CreER T2 ; Pten LoxP/LoxP ; Braf CA/+ mice or DNA tattoo administration of the pVAX1/mTyr-Cre construct on the flank of Pten LoxP/LoxP ; Braf CA/+ mice ( n = 5 per group). When the average tumor size was 25 mm 2 , tumor-bearing mice were placed on PLX4720 chow. Tumor outgrowth was monitored over time by digital photography, and tumor size was plotted against time from tumor induction for PLX4720-treated and control chow mock-treated animals. Data are expressed as the means ± SD.

    Journal: International Journal of Molecular Sciences

    Article Title: Dermal Delivery of Constructs Encoding Cre Recombinase to Induce Skin Tumors in PtenLoxP/LoxP;BrafCA/+ Mice

    doi: 10.3390/ijms17122149

    Figure Lengend Snippet: Inhibition of BRAF V600E in induced tumors led to tumor outgrowth inhibition. Melanomas were induced by 4-hydroxytamoxifen application on the flank of 4–10-week-old C57BL/6J Tyr::CreER T2 ; Pten LoxP/LoxP ; Braf CA/+ mice or DNA tattoo administration of the pVAX1/mTyr-Cre construct on the flank of Pten LoxP/LoxP ; Braf CA/+ mice ( n = 5 per group). When the average tumor size was 25 mm 2 , tumor-bearing mice were placed on PLX4720 chow. Tumor outgrowth was monitored over time by digital photography, and tumor size was plotted against time from tumor induction for PLX4720-treated and control chow mock-treated animals. Data are expressed as the means ± SD.

    Article Snippet: Plasmids for dermal naked DNA delivery were constructed by using the minimal pVAX1 vector (Life Technologies, Carlsbad, CA, USA) as a backbone. pVAX1/CAG-Cre was generated by the insertion of the CAG-Cre sequence from pCAG-Cre (a gift from Connie Cepko, Addgene Plasmid #13775) [ ] by restriction with SpeI/NotI, which resulted in the replacement of the CMV promoter previously present in the pVAX1 vector. pVAX1/noCMV-Cre was generated by first removing the CMV promoter in the pVAX1 backbone by Spe I digestion and direct re-ligation.

    Techniques: Inhibition, Mouse Assay, Construct

    S1 protein and N protein expression from the DNA vaccine in COS-7 cells. COS-7 cells were transfected with pVAX1-S1 (A), pVAX1-N (B), and backbone plasmid (C); 48 h later, protein expression was analyzed using an immunofluorescence assay.

    Journal: Clinical and Vaccine Immunology : CVI

    Article Title: Oral and Nasal DNA Vaccines Delivered by Attenuated Salmonella enterica Serovar Typhimurium Induce a Protective Immune Response against Infectious Bronchitis in Chickens ▿

    doi: 10.1128/CVI.00034-11

    Figure Lengend Snippet: S1 protein and N protein expression from the DNA vaccine in COS-7 cells. COS-7 cells were transfected with pVAX1-S1 (A), pVAX1-N (B), and backbone plasmid (C); 48 h later, protein expression was analyzed using an immunofluorescence assay.

    Article Snippet: The pVAX1 plasmid vector was purchased from Invitrogen and contained elements that complied with the Food and Drug Administration document Points to Consider on Plasmid DNA Vaccines for Preventative Infectious Disease Indications ( ).

    Techniques: Expressing, Transfection, Plasmid Preparation, Immunofluorescence

    Scheme of cloning GAPDH into vectors. (A) Cloning GAPDH into pMD-19T. (B) Cloning GAPDH into pVAX1. (C) Map of eukaryotic expression vector pVAX1.

    Journal: Frontiers in Microbiology

    Article Title: Protective Efficacy of Coccidial Common Antigen Glyceraldehyde 3-Phosphate Dehydrogenase (GAPDH) against Challenge with Three Eimeria Species

    doi: 10.3389/fmicb.2017.01245

    Figure Lengend Snippet: Scheme of cloning GAPDH into vectors. (A) Cloning GAPDH into pMD-19T. (B) Cloning GAPDH into pVAX1. (C) Map of eukaryotic expression vector pVAX1.

    Article Snippet: Plasmids, Parasites, and Animals The prokaryotic expression vector pET-32a was purchased from Novagen (Darmstadt, Germany), and the eukaryotic expression vector pVAX1 ( Figure ) was purchased from Invitrogen (Carlsbad, CA, United States).

    Techniques: Clone Assay, Expressing, Plasmid Preparation

    Figure 1. Construction and expression of esx constructs. ( A ) Schematic representation of all 5 trivalent esx constructs encompassing a total of 15 esx antigens. All genes are cloned into the pVAX1 mammalian vector and are under the CMV promoter.

    Journal: Human Vaccines & Immunotherapeutics

    Article Title: Multivalent TB vaccines targeting the esx gene family generate potent and broad cell-mediated immune responses superior to BCG

    doi: 10.4161/hv.29574

    Figure Lengend Snippet: Figure 1. Construction and expression of esx constructs. ( A ) Schematic representation of all 5 trivalent esx constructs encompassing a total of 15 esx antigens. All genes are cloned into the pVAX1 mammalian vector and are under the CMV promoter.

    Article Snippet: After obtaining the sequences each construct was commercially synthesized, species-specific codon and RNA optimized and subsequently subcloned (Genscript) into the pVAX1 mammalian expression vector (Invitrogen) under the control of the cytomegalovirus immediate-early promoter.

    Techniques: Expressing, Construct, Clone Assay, Plasmid Preparation

    Tumor regression after multiple deliveries of phIL-15 using electroporation. Tumor volume (mm 3 ) monitored over 50 days for non-responders ( A ) and responders ( B ) following intratumoral delivery of 50 pl of phIL-15 (2.0 mg/ml) on days 0, 3, and 6 using either six 100 μs pulses at 1300 V/cm or six 20 ms pulses at 500 V/cm delivered by penetrating electrode array. The group that received phIL-15 using six 20 ms pulses at 500 V/cm had one mouse represented on the non-responder graph (A), with a tumor that increased in volume up to day 30 and then regressed. On day 51 there was a measureable mass. Data is represented as the average percentage of the volume of the tumor at day zero. Error bars represent standard deviation. The number of responders or nonresponders is indicated on the graph as a fraction of the total number of animals in each group. P pulses; pVAX1 control plasmid; phIL-15 plasmid encoding the hIL-15 gene.

    Journal: Technology in Cancer Research & Treatment

    Article Title: Delivery of Interleukin-15 to B16 Melanoma by Electroporation Leads to Tumor Regression and Long-term Survival

    doi: 10.7785/tcrtexpress.2013.600252

    Figure Lengend Snippet: Tumor regression after multiple deliveries of phIL-15 using electroporation. Tumor volume (mm 3 ) monitored over 50 days for non-responders ( A ) and responders ( B ) following intratumoral delivery of 50 pl of phIL-15 (2.0 mg/ml) on days 0, 3, and 6 using either six 100 μs pulses at 1300 V/cm or six 20 ms pulses at 500 V/cm delivered by penetrating electrode array. The group that received phIL-15 using six 20 ms pulses at 500 V/cm had one mouse represented on the non-responder graph (A), with a tumor that increased in volume up to day 30 and then regressed. On day 51 there was a measureable mass. Data is represented as the average percentage of the volume of the tumor at day zero. Error bars represent standard deviation. The number of responders or nonresponders is indicated on the graph as a fraction of the total number of animals in each group. P pulses; pVAX1 control plasmid; phIL-15 plasmid encoding the hIL-15 gene.

    Article Snippet: The plasmid contains an optimized IL-15 sequence that was cloned into a pVAX1 cloning vector (Invitrogen, Carlsbad, CA) ( , ). phIL-15 and control vector pVAX1 were commercially prepared to endotoxin levels of < 100 EU/mg (Aldevron, Fargo, ND) and diluted in 0.9% sterile injectable saline to the appropriate concentration for each experiment.

    Techniques: Electroporation, Mass Spectrometry, Standard Deviation, Plasmid Preparation

    Increased T-cell proliferative potential following DNA vaccination plus co-administration of pRelA. Proliferative responses were measured seven days following the third vaccination with either pEnv or pGag alone, pEnv or pGag with pRelA molecular adjuvant, or empty vector control pVax1 alone. Splenocytes were incubated with recombinant HIV-1 Env ( A ) or Gag ( B ) at various concentrations: 0.5 (white bars), 1.0 (light gray bars), and 5.0 (dark gray bars) and subsequently pulsed with tritiated ( 3 H)-thymidine for 24 h. Incorporated thymidine was expressed as a stimulation index (SI) calculated by dividing the mean cpm (counts per minute) of Ag-stimulated wells by the mean cpm of non-stimulated wells. Fold increase in SI for pRelA-adjuvanted mice are displayed for each concentration of Env ( A , right panel) or Gag ( B , right panel). Samples were tested in triplicate. Error bars represent the SEM, and statistically significant values are provided for the indicated group comparison shown in the graphs. **** p

    Journal: Vaccines

    Article Title: Co-Administration of Molecular Adjuvants Expressing NF-Kappa B Subunit p65/RelA or Type-1 Transactivator T-bet Enhance Antigen Specific DNA Vaccine-Induced Immunity

    doi: 10.3390/vaccines2020196

    Figure Lengend Snippet: Increased T-cell proliferative potential following DNA vaccination plus co-administration of pRelA. Proliferative responses were measured seven days following the third vaccination with either pEnv or pGag alone, pEnv or pGag with pRelA molecular adjuvant, or empty vector control pVax1 alone. Splenocytes were incubated with recombinant HIV-1 Env ( A ) or Gag ( B ) at various concentrations: 0.5 (white bars), 1.0 (light gray bars), and 5.0 (dark gray bars) and subsequently pulsed with tritiated ( 3 H)-thymidine for 24 h. Incorporated thymidine was expressed as a stimulation index (SI) calculated by dividing the mean cpm (counts per minute) of Ag-stimulated wells by the mean cpm of non-stimulated wells. Fold increase in SI for pRelA-adjuvanted mice are displayed for each concentration of Env ( A , right panel) or Gag ( B , right panel). Samples were tested in triplicate. Error bars represent the SEM, and statistically significant values are provided for the indicated group comparison shown in the graphs. **** p

    Article Snippet: The optimized genes were then sub-cloned into modified pVax1 mammalian expression vectors (Invitrogen, Carlsbad, CA, USA) under the control of the cytomegalovirus immediate-early (CMV) promoter.

    Techniques: Plasmid Preparation, Incubation, Recombinant, Mouse Assay, Concentration Assay

    Transcription factor adjuvants enhance antigen specific DNA vaccine induced T cell immunity. ( A ) Balb/C mice ( n= 4 /group) were vaccinated three times at two week intervals with HIV-1 pGag or pEnv alone, pGag or pEnv with co delivery of either pRelA or pTbet. Other control groups were pRelA or pTbet alone, or a pVax1 control. T cell responses (CD8 + and CD4 + ) were analyzed by IFN-γ ELISPOT one week following the third immunization and results for IFN-γ+ spot forming cells (SFC) per 10 6 MACS-purified T cells are indicated following re-stimulation with subtype B HIV-1 Env ( B ) or Gag ( C ) peptide pools. Samples were performed in triplicate, error bars represent SEM, and statistically significant values are shown; ** p

    Journal: Vaccines

    Article Title: Co-Administration of Molecular Adjuvants Expressing NF-Kappa B Subunit p65/RelA or Type-1 Transactivator T-bet Enhance Antigen Specific DNA Vaccine-Induced Immunity

    doi: 10.3390/vaccines2020196

    Figure Lengend Snippet: Transcription factor adjuvants enhance antigen specific DNA vaccine induced T cell immunity. ( A ) Balb/C mice ( n= 4 /group) were vaccinated three times at two week intervals with HIV-1 pGag or pEnv alone, pGag or pEnv with co delivery of either pRelA or pTbet. Other control groups were pRelA or pTbet alone, or a pVax1 control. T cell responses (CD8 + and CD4 + ) were analyzed by IFN-γ ELISPOT one week following the third immunization and results for IFN-γ+ spot forming cells (SFC) per 10 6 MACS-purified T cells are indicated following re-stimulation with subtype B HIV-1 Env ( B ) or Gag ( C ) peptide pools. Samples were performed in triplicate, error bars represent SEM, and statistically significant values are shown; ** p

    Article Snippet: The optimized genes were then sub-cloned into modified pVax1 mammalian expression vectors (Invitrogen, Carlsbad, CA, USA) under the control of the cytomegalovirus immediate-early (CMV) promoter.

    Techniques: Mouse Assay, Enzyme-linked Immunospot, Magnetic Cell Separation, Purification

    Molecular adjuvant construction and expression. ( A ) Mouse RelA or T-bet primary sequences were genetically optimized, synthesized, and then subcloned into modified pVax1 expression vectors. Optimization entailed inclusion of a IgE leader peptide (IgE), preceded by a Kozak sequence, fused at the N -terminus. The figure indicates the restrictions enzymes used for subcloning, the translation initiation site (forward arrow), IgE leader peptide (IgE; hatched bar), protein length (aa), and transgenes (black with white lettering); ( B ) Protein expression from the nuclear extract was analyzed by Western immunoblotting following transfection of HEK 293T cells with pRelA, pTbet, or empty vector control (pVax1). The relative size (kDa) of the proteins are determined by detection analysis using protein-specific Abs as indicated; ( C ) Over expression of RelA potently induces κB dependent transcription. HeLa cells were transiently transfected with a NF-κB-dependent luciferase reporter gene together with expression vectors encoding RelA/p65. The cotransfected cells were subsequently grown for 48 h, and the luciferase activity was determined as described in the Materials and Methods; ( D ). Overexpression of T-bet stimulates production of IFN-γ: Naive CD4 T cells were transfected with either pT-bet or pVax1 and stimulated with anti-CD3 plus anti-CD28 followed the measurement of IFN-γ production by enzyme-linked immunosorbent assay (ELISA) as described Materials and Methods. IFN-γ levels are expressed as μg/mL

    Journal: Vaccines

    Article Title: Co-Administration of Molecular Adjuvants Expressing NF-Kappa B Subunit p65/RelA or Type-1 Transactivator T-bet Enhance Antigen Specific DNA Vaccine-Induced Immunity

    doi: 10.3390/vaccines2020196

    Figure Lengend Snippet: Molecular adjuvant construction and expression. ( A ) Mouse RelA or T-bet primary sequences were genetically optimized, synthesized, and then subcloned into modified pVax1 expression vectors. Optimization entailed inclusion of a IgE leader peptide (IgE), preceded by a Kozak sequence, fused at the N -terminus. The figure indicates the restrictions enzymes used for subcloning, the translation initiation site (forward arrow), IgE leader peptide (IgE; hatched bar), protein length (aa), and transgenes (black with white lettering); ( B ) Protein expression from the nuclear extract was analyzed by Western immunoblotting following transfection of HEK 293T cells with pRelA, pTbet, or empty vector control (pVax1). The relative size (kDa) of the proteins are determined by detection analysis using protein-specific Abs as indicated; ( C ) Over expression of RelA potently induces κB dependent transcription. HeLa cells were transiently transfected with a NF-κB-dependent luciferase reporter gene together with expression vectors encoding RelA/p65. The cotransfected cells were subsequently grown for 48 h, and the luciferase activity was determined as described in the Materials and Methods; ( D ). Overexpression of T-bet stimulates production of IFN-γ: Naive CD4 T cells were transfected with either pT-bet or pVax1 and stimulated with anti-CD3 plus anti-CD28 followed the measurement of IFN-γ production by enzyme-linked immunosorbent assay (ELISA) as described Materials and Methods. IFN-γ levels are expressed as μg/mL

    Article Snippet: The optimized genes were then sub-cloned into modified pVax1 mammalian expression vectors (Invitrogen, Carlsbad, CA, USA) under the control of the cytomegalovirus immediate-early (CMV) promoter.

    Techniques: Expressing, Synthesized, Modification, Sequencing, Subcloning, Western Blot, Transfection, Plasmid Preparation, Over Expression, Luciferase, Activity Assay, Enzyme-linked Immunosorbent Assay

    Schematic diagram of the construction of pVAX::HISAK70-asd and pVAX-asd. The kanamycin resistance gene in the eukaryotic expression vectors was replaced by the asd gene. a To generate pVAX::HISAK70 without kanamycin resistance cassette (Kan r ), inverse PCR with primers noKan Pac I-1R 5’-CTTGTTTAATTAA GCGAAACGATCCTCATCCTGTC-3’ ( Pac I site underlined) and noKan Kpn I-1D 5’-GACGAGGGTACCATTATTAACGCTTACAATTTC-3’ ( Kpn I site underlined) were employed for outward amplification. The asd gene starting at bp 170 and ending at bp 1345 was PCR amplified from bacterial chromosomal with primers asd Kpn I-1D 5’ CTGCAAGGTACCCTACGCCAACTGGCGCAGCAT-3’ ( Kpn I site underlined) and the asd Pac I-1R 5’-TTGGCTGTTAATTAAATGGTGAAGGATGCGCCACAG-3’ ( Pac I site underlined) creating an amplicon with Pac I and Kpn I sites on its ends. b To generate pVAX1 without Kan r , inverse PCR with primers noKan Pac I-1R and noKan Kpn I-1D were employed for outward amplification. c – d The PCR products of asd , ∆kan r pVAX1::HISAK70 and ∆kan r pVAX1 were each double digested with Pac I and Kpn I, gel isolated, ligated, and transformed. Thus, pVAX1::HisAK70-asd and pVAX1-asd without resistance gene were obtained

    Journal: Parasites & Vectors

    Article Title: HisAK70: progress towards a vaccine against different forms of leishmaniosis

    doi: 10.1186/s13071-015-1246-y

    Figure Lengend Snippet: Schematic diagram of the construction of pVAX::HISAK70-asd and pVAX-asd. The kanamycin resistance gene in the eukaryotic expression vectors was replaced by the asd gene. a To generate pVAX::HISAK70 without kanamycin resistance cassette (Kan r ), inverse PCR with primers noKan Pac I-1R 5’-CTTGTTTAATTAA GCGAAACGATCCTCATCCTGTC-3’ ( Pac I site underlined) and noKan Kpn I-1D 5’-GACGAGGGTACCATTATTAACGCTTACAATTTC-3’ ( Kpn I site underlined) were employed for outward amplification. The asd gene starting at bp 170 and ending at bp 1345 was PCR amplified from bacterial chromosomal with primers asd Kpn I-1D 5’ CTGCAAGGTACCCTACGCCAACTGGCGCAGCAT-3’ ( Kpn I site underlined) and the asd Pac I-1R 5’-TTGGCTGTTAATTAAATGGTGAAGGATGCGCCACAG-3’ ( Pac I site underlined) creating an amplicon with Pac I and Kpn I sites on its ends. b To generate pVAX1 without Kan r , inverse PCR with primers noKan Pac I-1R and noKan Kpn I-1D were employed for outward amplification. c – d The PCR products of asd , ∆kan r pVAX1::HISAK70 and ∆kan r pVAX1 were each double digested with Pac I and Kpn I, gel isolated, ligated, and transformed. Thus, pVAX1::HisAK70-asd and pVAX1-asd without resistance gene were obtained

    Article Snippet: This expression cassette was cloned into the eukaryotic expression plasmid pVAX1 (Invitrogen) to obtain the recombinant plasmid pVAX1::HisAK70 .

    Techniques: Expressing, Inverse PCR, Amplification, Polymerase Chain Reaction, Isolation, Transformation Assay

    Salivary PAc-specific IgA and serum PAc-specific IgG antibody levels in rats immunized intranasally with pCIA-P in combination with pCCL17/VAX, pCCL19/VAX, pCCL17/VAX plus pCCL19/VAX, or pVAX1. Saliva and serum samples were collected 52 days after the first immunization. The specific anti-PAc salivary IgA (A) and specific anti-PAc serum IgG (B) concentrations were determined by ELISA. The data are expressed as the mean±SD. n =5. * P

    Journal: Acta Pharmacologica Sinica

    Article Title: CCL17 combined with CCL19 as a nasal adjuvant enhances the immunogenicity of an anti-caries DNA vaccine in rodents

    doi: 10.1038/aps.2016.73

    Figure Lengend Snippet: Salivary PAc-specific IgA and serum PAc-specific IgG antibody levels in rats immunized intranasally with pCIA-P in combination with pCCL17/VAX, pCCL19/VAX, pCCL17/VAX plus pCCL19/VAX, or pVAX1. Saliva and serum samples were collected 52 days after the first immunization. The specific anti-PAc salivary IgA (A) and specific anti-PAc serum IgG (B) concentrations were determined by ELISA. The data are expressed as the mean±SD. n =5. * P

    Article Snippet: Plasmid construction The murine CCL19 and CCL17 genes were amplified from mouse spleen by RT-PCR and cloned into the Nhe I and Kpn I sites of the pVAX1 vector (Clontech Laboratories, Inc, Mountain View, CA, USA) to obtain plasmids encoding CCL19 and CCL17, which were designated pCCL19/pVAX and pCCL17/pVAX, respectively.

    Techniques: Enzyme-linked Immunosorbent Assay

    Caries scores of rats immunized intranasally with pCIA-P in combination with pCCL17/VAX, pCCL19/VAX, pCCL17/VAX plus pCCL19/VAX, or pVAX1. ** P

    Journal: Acta Pharmacologica Sinica

    Article Title: CCL17 combined with CCL19 as a nasal adjuvant enhances the immunogenicity of an anti-caries DNA vaccine in rodents

    doi: 10.1038/aps.2016.73

    Figure Lengend Snippet: Caries scores of rats immunized intranasally with pCIA-P in combination with pCCL17/VAX, pCCL19/VAX, pCCL17/VAX plus pCCL19/VAX, or pVAX1. ** P

    Article Snippet: Plasmid construction The murine CCL19 and CCL17 genes were amplified from mouse spleen by RT-PCR and cloned into the Nhe I and Kpn I sites of the pVAX1 vector (Clontech Laboratories, Inc, Mountain View, CA, USA) to obtain plasmids encoding CCL19 and CCL17, which were designated pCCL19/pVAX and pCCL17/pVAX, respectively.

    Techniques:

    CCL17 and CCL19 increase the number of mature DCs in secondary lymphoid tissues. Splenocytes and DLN cells were collected from mice treated with pCCL17/VAX, pCCL19/VAX, pCCL17/VAX plus pCCL19/VAX, or pVAX1, digested with collagenase IV 3 days after immunization, stained with a DC marker (CD11c) and a class II marker (I-Ab α chain), and analyzed by flow cytometry. This result is representative of three experiments.

    Journal: Acta Pharmacologica Sinica

    Article Title: CCL17 combined with CCL19 as a nasal adjuvant enhances the immunogenicity of an anti-caries DNA vaccine in rodents

    doi: 10.1038/aps.2016.73

    Figure Lengend Snippet: CCL17 and CCL19 increase the number of mature DCs in secondary lymphoid tissues. Splenocytes and DLN cells were collected from mice treated with pCCL17/VAX, pCCL19/VAX, pCCL17/VAX plus pCCL19/VAX, or pVAX1, digested with collagenase IV 3 days after immunization, stained with a DC marker (CD11c) and a class II marker (I-Ab α chain), and analyzed by flow cytometry. This result is representative of three experiments.

    Article Snippet: Plasmid construction The murine CCL19 and CCL17 genes were amplified from mouse spleen by RT-PCR and cloned into the Nhe I and Kpn I sites of the pVAX1 vector (Clontech Laboratories, Inc, Mountain View, CA, USA) to obtain plasmids encoding CCL19 and CCL17, which were designated pCCL19/pVAX and pCCL17/pVAX, respectively.

    Techniques: Mouse Assay, Staining, Marker, Flow Cytometry, Cytometry

    Salivary PAc-specific IgA and serum PAc-specific IgG antibody levels in mice immunized intranasally with pCIA-P in combination with pCCL17/VAX, pCCL19/VAX, pCCL17/VAX plus pCCL19/VAX, or pVAX1. Saliva and serum samples were collected at 4, 6, 8, 10, 12, 14, and 16 weeks after the first immunization. The specific anti-PAc salivary IgA (A) and specific anti-PAc serum IgG (B) concentrations were determined by ELISA. The data are expressed as the mean±SD. n =5. * P

    Journal: Acta Pharmacologica Sinica

    Article Title: CCL17 combined with CCL19 as a nasal adjuvant enhances the immunogenicity of an anti-caries DNA vaccine in rodents

    doi: 10.1038/aps.2016.73

    Figure Lengend Snippet: Salivary PAc-specific IgA and serum PAc-specific IgG antibody levels in mice immunized intranasally with pCIA-P in combination with pCCL17/VAX, pCCL19/VAX, pCCL17/VAX plus pCCL19/VAX, or pVAX1. Saliva and serum samples were collected at 4, 6, 8, 10, 12, 14, and 16 weeks after the first immunization. The specific anti-PAc salivary IgA (A) and specific anti-PAc serum IgG (B) concentrations were determined by ELISA. The data are expressed as the mean±SD. n =5. * P

    Article Snippet: Plasmid construction The murine CCL19 and CCL17 genes were amplified from mouse spleen by RT-PCR and cloned into the Nhe I and Kpn I sites of the pVAX1 vector (Clontech Laboratories, Inc, Mountain View, CA, USA) to obtain plasmids encoding CCL19 and CCL17, which were designated pCCL19/pVAX and pCCL17/pVAX, respectively.

    Techniques: Mouse Assay, Enzyme-linked Immunosorbent Assay

    Restriction Enzyme Analysis of Recombinant pVAX1-SAG1 Line 1 and 2, the recombinant pVAX-SAG1 plasmid digested by NheI; Line 3, 1 kb DNA ladder; Line 4, the pVAX vector digested by NheI.

    Journal: Jundishapur Journal of Microbiology

    Article Title: Cloning and Expression of Major Surface Antigen 1 Gene of Toxoplasma gondii RH Strain Using the Expression Vector pVAX1 in Chinese Hamster Ovary Cells

    doi: 10.5812/jjm.22570

    Figure Lengend Snippet: Restriction Enzyme Analysis of Recombinant pVAX1-SAG1 Line 1 and 2, the recombinant pVAX-SAG1 plasmid digested by NheI; Line 3, 1 kb DNA ladder; Line 4, the pVAX vector digested by NheI.

    Article Snippet: The Escherichia coli strain DH5α and pVAX1 expression vector were purchased from Novagen (Novagen Inc., Madison, WI, USA).

    Techniques: Recombinant, Plasmid Preparation

    Expression Analysis of SAG1 in CHO Cells by Immunofluorescence Assay A) Transfected CHO cells with empty pVAX1 plasmid; B) DAPI staining of negative control group; C) Transfected CHO cells with pVAX1-SAG1 and revealed using anti-SAG1 monoclonal antibody and secondary antibody labeled with FITC; D) DAPI was used to stain the cell nuclei.

    Journal: Jundishapur Journal of Microbiology

    Article Title: Cloning and Expression of Major Surface Antigen 1 Gene of Toxoplasma gondii RH Strain Using the Expression Vector pVAX1 in Chinese Hamster Ovary Cells

    doi: 10.5812/jjm.22570

    Figure Lengend Snippet: Expression Analysis of SAG1 in CHO Cells by Immunofluorescence Assay A) Transfected CHO cells with empty pVAX1 plasmid; B) DAPI staining of negative control group; C) Transfected CHO cells with pVAX1-SAG1 and revealed using anti-SAG1 monoclonal antibody and secondary antibody labeled with FITC; D) DAPI was used to stain the cell nuclei.

    Article Snippet: The Escherichia coli strain DH5α and pVAX1 expression vector were purchased from Novagen (Novagen Inc., Madison, WI, USA).

    Techniques: Expressing, Immunofluorescence, Transfection, Plasmid Preparation, Staining, Negative Control, Labeling

    Restriction Enzyme Analysis of Recombinant pVAX1-SAG1 Line 1 and 2, the recombinant pVAX-SAG1 plasmid digested by NheI and XhoI; Line M, 1 kb DNA ladder.

    Journal: Jundishapur Journal of Microbiology

    Article Title: Cloning and Expression of Major Surface Antigen 1 Gene of Toxoplasma gondii RH Strain Using the Expression Vector pVAX1 in Chinese Hamster Ovary Cells

    doi: 10.5812/jjm.22570

    Figure Lengend Snippet: Restriction Enzyme Analysis of Recombinant pVAX1-SAG1 Line 1 and 2, the recombinant pVAX-SAG1 plasmid digested by NheI and XhoI; Line M, 1 kb DNA ladder.

    Article Snippet: The Escherichia coli strain DH5α and pVAX1 expression vector were purchased from Novagen (Novagen Inc., Madison, WI, USA).

    Techniques: Recombinant, Plasmid Preparation

    Agarose Gel Electrophoresis Analysis the Expression of SAG1 in CHO Cells by RT-PCR Assay Line M, 1 kb DNA ladder; Line 1, Transfected CHO cells with empty pVAX1 plasmid; Line 2, Transfected CHO cells with pVAX-SAG1.

    Journal: Jundishapur Journal of Microbiology

    Article Title: Cloning and Expression of Major Surface Antigen 1 Gene of Toxoplasma gondii RH Strain Using the Expression Vector pVAX1 in Chinese Hamster Ovary Cells

    doi: 10.5812/jjm.22570

    Figure Lengend Snippet: Agarose Gel Electrophoresis Analysis the Expression of SAG1 in CHO Cells by RT-PCR Assay Line M, 1 kb DNA ladder; Line 1, Transfected CHO cells with empty pVAX1 plasmid; Line 2, Transfected CHO cells with pVAX-SAG1.

    Article Snippet: The Escherichia coli strain DH5α and pVAX1 expression vector were purchased from Novagen (Novagen Inc., Madison, WI, USA).

    Techniques: Agarose Gel Electrophoresis, Expressing, Reverse Transcription Polymerase Chain Reaction, Transfection, Plasmid Preparation

    Expression of SAG1 Protein in Transfected CHO Cell Lysate by Western Blot Lane M, Prestained Protein Ladder; Lane 1, untransfected CHO cells; Lane 2, transfected CHO cells with empty pVAX1 plasmid; Lane 3, transfected CHO cells with pVAX1-SAG1 expression plasmid; a protein band about 30 kDa corresponding to SAG1 protein is detected.

    Journal: Jundishapur Journal of Microbiology

    Article Title: Cloning and Expression of Major Surface Antigen 1 Gene of Toxoplasma gondii RH Strain Using the Expression Vector pVAX1 in Chinese Hamster Ovary Cells

    doi: 10.5812/jjm.22570

    Figure Lengend Snippet: Expression of SAG1 Protein in Transfected CHO Cell Lysate by Western Blot Lane M, Prestained Protein Ladder; Lane 1, untransfected CHO cells; Lane 2, transfected CHO cells with empty pVAX1 plasmid; Lane 3, transfected CHO cells with pVAX1-SAG1 expression plasmid; a protein band about 30 kDa corresponding to SAG1 protein is detected.

    Article Snippet: The Escherichia coli strain DH5α and pVAX1 expression vector were purchased from Novagen (Novagen Inc., Madison, WI, USA).

    Techniques: Expressing, Transfection, Western Blot, Plasmid Preparation

    Agarose Gel Electrophoresis Analysis of Purified pVAX-SAG1 Recombinant Plasmids From Five Colonies on 1% Agarose Gel Line. M, 1 kb DNA ladder; Lane 1, negative control; Lane 2 - 6: selected pVAX1-SAG1 recombinant plasmids.

    Journal: Jundishapur Journal of Microbiology

    Article Title: Cloning and Expression of Major Surface Antigen 1 Gene of Toxoplasma gondii RH Strain Using the Expression Vector pVAX1 in Chinese Hamster Ovary Cells

    doi: 10.5812/jjm.22570

    Figure Lengend Snippet: Agarose Gel Electrophoresis Analysis of Purified pVAX-SAG1 Recombinant Plasmids From Five Colonies on 1% Agarose Gel Line. M, 1 kb DNA ladder; Lane 1, negative control; Lane 2 - 6: selected pVAX1-SAG1 recombinant plasmids.

    Article Snippet: The Escherichia coli strain DH5α and pVAX1 expression vector were purchased from Novagen (Novagen Inc., Madison, WI, USA).

    Techniques: Agarose Gel Electrophoresis, Purification, Recombinant, Negative Control