plvx ires zsgreen1  (TaKaRa)

 
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    Name:
    pLVX IRES ZsGreen1 Vector
    Description:
    Living Colors ZsGreen1 is an exceptionally bright green fluorescent protein derived from a Zoanthus sp reef coral Matz et al 1999 that has been modified for high solubility bright emission and rapid chromophore maturation ZsGreen1 is the brightest commercially available green fluorescent protein up to 4X brighter than EGFP and is ideally suited for whole cell labelling promoter reporter studies or as a transfection control
    Catalog Number:
    632187
    Price:
    None
    Size:
    10 ug
    Category:
    ZsGreen1 fluorescent protein Cyan and green fluorescent proteins Fluorescent protein plasmids Fluorescent proteins Gene function
    Buy from Supplier


    Structured Review

    TaKaRa plvx ires zsgreen1
    ORF2 and ORF3 are required for releasing viral particles to infect naïve HepG2C3A cells. ( A ) Schematic representation of the transcomplementation system for packaging HEV virions. ( B ) Representative flow cytometry plots demonstrating efficient ORF2 and ORF3 expression. HepG2C3A cells were transduced with <t>pLVX-ORF2-IRES-zsGreen</t> and/or pLEX-ORF3-IRES-mCherry or not transduced. Flow cytometric analysis was performed 4 d following transduction to quantify the frequencies of ORF2 and/or ORF3 expressing cells. ( C ) Infection kinetics of transcomplemented HEV in HepG2C3A cells. Cell culture supernatants from naïve HepG2C3A, or HepG2C3A cells transduced with HEV ORF2 or/and ORF3, were collected 5 d posttransfection with rHEVΔORF2/3[Gluc] RNA. Naïve HepG2C3A cells were incubated with these supernatants. After 12 h, cells were washed and Gaussia luciferase activity quantified in the cell culture supernatants at the indicated time points. ( D ) Five days following transfection of rHEVΔORF2/3[Gluc] RNA into HepG2C3A, or HepG2C3A cells transduced with HEV ORF2 or/and ORF3, lysates were used to infect naïve HepG2C3A cells. Gluc activity was measured in the supernatants 4 d postinfection. Shown are averages and SDs of triplicate measurements of three independent experiments. * P
    Living Colors ZsGreen1 is an exceptionally bright green fluorescent protein derived from a Zoanthus sp reef coral Matz et al 1999 that has been modified for high solubility bright emission and rapid chromophore maturation ZsGreen1 is the brightest commercially available green fluorescent protein up to 4X brighter than EGFP and is ideally suited for whole cell labelling promoter reporter studies or as a transfection control
    https://www.bioz.com/result/plvx ires zsgreen1/product/TaKaRa
    Average 99 stars, based on 10 article reviews
    Price from $9.99 to $1999.99
    plvx ires zsgreen1 - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "Hepatitis E virus ORF3 is a functional ion channel required for release of infectious particles"

    Article Title: Hepatitis E virus ORF3 is a functional ion channel required for release of infectious particles

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi: 10.1073/pnas.1614955114

    ORF2 and ORF3 are required for releasing viral particles to infect naïve HepG2C3A cells. ( A ) Schematic representation of the transcomplementation system for packaging HEV virions. ( B ) Representative flow cytometry plots demonstrating efficient ORF2 and ORF3 expression. HepG2C3A cells were transduced with pLVX-ORF2-IRES-zsGreen and/or pLEX-ORF3-IRES-mCherry or not transduced. Flow cytometric analysis was performed 4 d following transduction to quantify the frequencies of ORF2 and/or ORF3 expressing cells. ( C ) Infection kinetics of transcomplemented HEV in HepG2C3A cells. Cell culture supernatants from naïve HepG2C3A, or HepG2C3A cells transduced with HEV ORF2 or/and ORF3, were collected 5 d posttransfection with rHEVΔORF2/3[Gluc] RNA. Naïve HepG2C3A cells were incubated with these supernatants. After 12 h, cells were washed and Gaussia luciferase activity quantified in the cell culture supernatants at the indicated time points. ( D ) Five days following transfection of rHEVΔORF2/3[Gluc] RNA into HepG2C3A, or HepG2C3A cells transduced with HEV ORF2 or/and ORF3, lysates were used to infect naïve HepG2C3A cells. Gluc activity was measured in the supernatants 4 d postinfection. Shown are averages and SDs of triplicate measurements of three independent experiments. * P
    Figure Legend Snippet: ORF2 and ORF3 are required for releasing viral particles to infect naïve HepG2C3A cells. ( A ) Schematic representation of the transcomplementation system for packaging HEV virions. ( B ) Representative flow cytometry plots demonstrating efficient ORF2 and ORF3 expression. HepG2C3A cells were transduced with pLVX-ORF2-IRES-zsGreen and/or pLEX-ORF3-IRES-mCherry or not transduced. Flow cytometric analysis was performed 4 d following transduction to quantify the frequencies of ORF2 and/or ORF3 expressing cells. ( C ) Infection kinetics of transcomplemented HEV in HepG2C3A cells. Cell culture supernatants from naïve HepG2C3A, or HepG2C3A cells transduced with HEV ORF2 or/and ORF3, were collected 5 d posttransfection with rHEVΔORF2/3[Gluc] RNA. Naïve HepG2C3A cells were incubated with these supernatants. After 12 h, cells were washed and Gaussia luciferase activity quantified in the cell culture supernatants at the indicated time points. ( D ) Five days following transfection of rHEVΔORF2/3[Gluc] RNA into HepG2C3A, or HepG2C3A cells transduced with HEV ORF2 or/and ORF3, lysates were used to infect naïve HepG2C3A cells. Gluc activity was measured in the supernatants 4 d postinfection. Shown are averages and SDs of triplicate measurements of three independent experiments. * P

    Techniques Used: Flow Cytometry, Cytometry, Expressing, Transduction, Infection, Cell Culture, Incubation, Luciferase, Activity Assay, Transfection

    2) Product Images from "Selection of an optimal promoter for gene transfer in normal B cells"

    Article Title: Selection of an optimal promoter for gene transfer in normal B cells

    Journal: Molecular Medicine Reports

    doi: 10.3892/mmr.2017.6974

    Transduction of Raji cells with plasmids containing various promoters. Raji lymphoma cells were modified with pLVX ZsGreen1, pLVTHM, pGIPZ and HIV-SFFV-RFP. The percentage of fluorescent (GFP- or RFP-positive) cells was determined using flow cytometry. GFP, green fluorescent protein; RFP, red fluorescent protein; HIV, human immunodeficiency virus; SFFV, spleen focus-forming virus.
    Figure Legend Snippet: Transduction of Raji cells with plasmids containing various promoters. Raji lymphoma cells were modified with pLVX ZsGreen1, pLVTHM, pGIPZ and HIV-SFFV-RFP. The percentage of fluorescent (GFP- or RFP-positive) cells was determined using flow cytometry. GFP, green fluorescent protein; RFP, red fluorescent protein; HIV, human immunodeficiency virus; SFFV, spleen focus-forming virus.

    Techniques Used: Transduction, Modification, Flow Cytometry, Cytometry

    3) Product Images from "Selection of an optimal promoter for gene transfer in normal B cells"

    Article Title: Selection of an optimal promoter for gene transfer in normal B cells

    Journal: Molecular Medicine Reports

    doi: 10.3892/mmr.2017.6974

    Transduction of Raji cells with plasmids containing various promoters. Raji lymphoma cells were modified with pLVX ZsGreen1, pLVTHM, pGIPZ and HIV-SFFV-RFP. The percentage of fluorescent (GFP- or RFP-positive) cells was determined using flow cytometry.
    Figure Legend Snippet: Transduction of Raji cells with plasmids containing various promoters. Raji lymphoma cells were modified with pLVX ZsGreen1, pLVTHM, pGIPZ and HIV-SFFV-RFP. The percentage of fluorescent (GFP- or RFP-positive) cells was determined using flow cytometry.

    Techniques Used: Transduction, Modification, Flow Cytometry, Cytometry

    4) Product Images from "Selection of an optimal promoter for gene transfer in normal B cells"

    Article Title: Selection of an optimal promoter for gene transfer in normal B cells

    Journal: Molecular Medicine Reports

    doi: 10.3892/mmr.2017.6974

    Transduction of Raji cells with plasmids containing various promoters. Raji lymphoma cells were modified with pLVX ZsGreen1, pLVTHM, pGIPZ and HIV-SFFV-RFP. The percentage of fluorescent (GFP- or RFP-positive) cells was determined using flow cytometry. GFP, green fluorescent protein; RFP, red fluorescent protein; HIV, human immunodeficiency virus; SFFV, spleen focus-forming virus.
    Figure Legend Snippet: Transduction of Raji cells with plasmids containing various promoters. Raji lymphoma cells were modified with pLVX ZsGreen1, pLVTHM, pGIPZ and HIV-SFFV-RFP. The percentage of fluorescent (GFP- or RFP-positive) cells was determined using flow cytometry. GFP, green fluorescent protein; RFP, red fluorescent protein; HIV, human immunodeficiency virus; SFFV, spleen focus-forming virus.

    Techniques Used: Transduction, Modification, Flow Cytometry, Cytometry

    5) Product Images from "Folate receptor 1 (FOLR1) targeted chimeric antigen receptor (CAR) T cells for the treatment of gastric cancer"

    Article Title: Folate receptor 1 (FOLR1) targeted chimeric antigen receptor (CAR) T cells for the treatment of gastric cancer

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0198347

    Generation and characterization of FOLR1-CAR T cells. T cells were obtained from human PBMCs of three healthy donors and the FOLR1-CAR gene was cloned into the pLVX-IRES-GFP vector and the lentivirus infection efficiency was determined. (A) The growth rate of infected or untransduced T cells was measured by CellTiter-Glo assay for 12 days. (B) Lentivirus infection rate was analyzed by GFP fluorescence on day 12 using FACS analysis. (C) Expression of FOLR1-CAR in T cells was measured by western blot analysis. (D-E) The population of T cells was evaluated by FACS analysis using CD3, CD4, and CD8 antibodies on day 12. Experiments were repeated three times with similar results. The data are represented as the mean of luminescence ± SD and positive rate ± SD (%) from triplicate cultures. Statistical analysis was performed using the two-sided unpaired t-test.
    Figure Legend Snippet: Generation and characterization of FOLR1-CAR T cells. T cells were obtained from human PBMCs of three healthy donors and the FOLR1-CAR gene was cloned into the pLVX-IRES-GFP vector and the lentivirus infection efficiency was determined. (A) The growth rate of infected or untransduced T cells was measured by CellTiter-Glo assay for 12 days. (B) Lentivirus infection rate was analyzed by GFP fluorescence on day 12 using FACS analysis. (C) Expression of FOLR1-CAR in T cells was measured by western blot analysis. (D-E) The population of T cells was evaluated by FACS analysis using CD3, CD4, and CD8 antibodies on day 12. Experiments were repeated three times with similar results. The data are represented as the mean of luminescence ± SD and positive rate ± SD (%) from triplicate cultures. Statistical analysis was performed using the two-sided unpaired t-test.

    Techniques Used: Clone Assay, Plasmid Preparation, Infection, Glo Assay, Fluorescence, FACS, Expressing, Western Blot

    6) Product Images from "Selection of an optimal promoter for gene transfer in normal B cells"

    Article Title: Selection of an optimal promoter for gene transfer in normal B cells

    Journal: Molecular Medicine Reports

    doi: 10.3892/mmr.2017.6974

    Transduction of Raji cells with plasmids containing various promoters. Raji lymphoma cells were modified with pLVX ZsGreen1, pLVTHM, pGIPZ and HIV-SFFV-RFP. The percentage of fluorescent (GFP- or RFP-positive) cells was determined using flow cytometry.
    Figure Legend Snippet: Transduction of Raji cells with plasmids containing various promoters. Raji lymphoma cells were modified with pLVX ZsGreen1, pLVTHM, pGIPZ and HIV-SFFV-RFP. The percentage of fluorescent (GFP- or RFP-positive) cells was determined using flow cytometry.

    Techniques Used: Transduction, Modification, Flow Cytometry, Cytometry

    7) Product Images from "Folate receptor 1 (FOLR1) targeted chimeric antigen receptor (CAR) T cells for the treatment of gastric cancer"

    Article Title: Folate receptor 1 (FOLR1) targeted chimeric antigen receptor (CAR) T cells for the treatment of gastric cancer

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0198347

    Generation and characterization of FOLR1-CAR T cells. T cells were obtained from human PBMCs of three healthy donors and the FOLR1-CAR gene was cloned into the pLVX-IRES-GFP vector and the lentivirus infection efficiency was determined. (A) The growth rate of infected or untransduced T cells was measured by CellTiter-Glo assay for 12 days. (B) Lentivirus infection rate was analyzed by GFP fluorescence on day 12 using FACS analysis. (C) Expression of FOLR1-CAR in T cells was measured by western blot analysis. (D-E) The population of T cells was evaluated by FACS analysis using CD3, CD4, and CD8 antibodies on day 12. Experiments were repeated three times with similar results. The data are represented as the mean of luminescence ± SD and positive rate ± SD (%) from triplicate cultures. Statistical analysis was performed using the two-sided unpaired t-test.
    Figure Legend Snippet: Generation and characterization of FOLR1-CAR T cells. T cells were obtained from human PBMCs of three healthy donors and the FOLR1-CAR gene was cloned into the pLVX-IRES-GFP vector and the lentivirus infection efficiency was determined. (A) The growth rate of infected or untransduced T cells was measured by CellTiter-Glo assay for 12 days. (B) Lentivirus infection rate was analyzed by GFP fluorescence on day 12 using FACS analysis. (C) Expression of FOLR1-CAR in T cells was measured by western blot analysis. (D-E) The population of T cells was evaluated by FACS analysis using CD3, CD4, and CD8 antibodies on day 12. Experiments were repeated three times with similar results. The data are represented as the mean of luminescence ± SD and positive rate ± SD (%) from triplicate cultures. Statistical analysis was performed using the two-sided unpaired t-test.

    Techniques Used: Clone Assay, Plasmid Preparation, Infection, Glo Assay, Fluorescence, FACS, Expressing, Western Blot

    Related Articles

    Transduction:

    Article Title: A novel miR-375-HOXB3-CDCA3/DNMT3B regulatory circuitry contributes to leukemogenesis in acute myeloid leukemia
    Article Snippet: .. Engraftment of NOD/SCID-IL2Rγ mice (NSG) THP1 cells were transduced with lentivirus vector pLVX-IRES-ZsGreen1 (Clontech), followed by sorting to obtain GFP-positive cells. .. GFP-positive THP1 cells were transduced with MSCV-miR-375 or MSCV-NC, followed by puromycin selection for 1 week.

    Article Title: Selection of an optimal promoter for gene transfer in normal B cells
    Article Snippet: .. B cells activated with CD40L and IL-21 are resistant to transduction To determine the levels of transgene expression in B cells three bicistronic plasmids were used that encode various GFPs under the control of different promoters, namely pGIPZ [cytomegalovirus (CMV) promoter; turbo GFP, which is an improved variant of the green fluorescent protein CopGFP], pLVTHM [elongation factor 1 alpha (EF1α) promoter; GFP) and pLVX-IRES-ZsGreen1 (CMV promoter; ZsGreen1, which is a human codon-optimized variant of ZsGreen) ( ). .. Independent of the vectors used in the present study, a very low level of transgene expression was detected in B cells; expression did not exceed 10%, as assessed with flow cytometry 7 days post-transduction ( ).

    Clone Assay:

    Article Title: Hepatitis E virus ORF3 is a functional ion channel required for release of infectious particles
    Article Snippet: .. To construct a lentiviral construct that encodes Kernow C1/p6 ORF2 or Kernow C1/p6 ORF3, the Kernow C1/p6 ORF2 or Kernow C1/p6 ORF3 cDNA was amplified by PCR from the HEV Kernow C1/p6 construct (a kind gift from Suzanne Emerson, NIH, Bethesda, MD) and then cloned into pLVX-IRES-zsGreen1 or pLEX-IRES-mCherry vectors using the In-Fusion HD Cloning Kit (Clontech). .. To construct the pLEX-IAV M2-IRES-mCherry vector, cDNA encoding Influenza A virus M2 (A/Puerto Rico/8/34/Mount Sinai/Wi(H1N1)) was synthesized by IDT with gBlock, and then cloned into pLEX-IRES-mCherry vectors using In-Fusion HD Cloning Kit (Clontech).

    Article Title: A novel miR-375-HOXB3-CDCA3/DNMT3B regulatory circuitry contributes to leukemogenesis in acute myeloid leukemia
    Article Snippet: .. To produce plasmids expressing HOXB3 and DNMT3B, human HOXB3 and DNMT3B coding sequences were amplified by PCR and then cloned into lentivirus vector pLVX-IRES-ZsGreen1 (Clontech) and pMSCV-puro (Clontech), respectively. .. To construct pMIR-HOXB3 3′UTR plasmid, human HOXB3 3′UTR was amplified by PCR and cloned into pMIR-REPORT vector (Ambion, Dallas, TX, USA).

    Article Title: A novel miR-375-HOXB3-CDCA3/DNMT3B regulatory circuitry contributes to leukemogenesis in acute myeloid leukemia
    Article Snippet: .. To produce plasmids expressing HOXB3 and DNMT3B, human HOXB3 and DNMT3B coding sequences were amplified by PCR and then cloned into lentivirus vector pLVX-IRES-ZsGreen1 (Clontech) and pMSCV-puro (Clontech), respectively. .. To construct pMIR-HOXB3 3′UTR plasmid, human HOXB3 3′UTR was amplified by PCR and cloned into pMIR-REPORT vector (Ambion, Dallas, TX, USA).

    Article Title: Identification of the Intragenomic Promoter Controlling Hepatitis E Virus Subgenomic RNA Transcription
    Article Snippet: .. To construct lentiviral constructs encoding ORF1 of Kernow C1/p6 (GenBank accession number JQ679013 ), the Kernow C1/p6 ORF1 cDNA was amplified by PCR from a plasmid encoding the full-length (FL) infectious HEV clone Kernow C1/p6 (a kind gift from Suzanne Emerson, NIH) and then cloned into pLVX-IRES-zsGreen1 vector using an In-Fusion HD cloning kit (Clontech, Mountain View, CA, USA). .. The GAD mutant of ORF1 inactivating the polymerase was generated by QuikChange (Stratagene) site-directed mutagenesis.

    Amplification:

    Article Title: Hepatitis E virus ORF3 is a functional ion channel required for release of infectious particles
    Article Snippet: .. To construct a lentiviral construct that encodes Kernow C1/p6 ORF2 or Kernow C1/p6 ORF3, the Kernow C1/p6 ORF2 or Kernow C1/p6 ORF3 cDNA was amplified by PCR from the HEV Kernow C1/p6 construct (a kind gift from Suzanne Emerson, NIH, Bethesda, MD) and then cloned into pLVX-IRES-zsGreen1 or pLEX-IRES-mCherry vectors using the In-Fusion HD Cloning Kit (Clontech). .. To construct the pLEX-IAV M2-IRES-mCherry vector, cDNA encoding Influenza A virus M2 (A/Puerto Rico/8/34/Mount Sinai/Wi(H1N1)) was synthesized by IDT with gBlock, and then cloned into pLEX-IRES-mCherry vectors using In-Fusion HD Cloning Kit (Clontech).

    Article Title: A novel miR-375-HOXB3-CDCA3/DNMT3B regulatory circuitry contributes to leukemogenesis in acute myeloid leukemia
    Article Snippet: .. To produce plasmids expressing HOXB3 and DNMT3B, human HOXB3 and DNMT3B coding sequences were amplified by PCR and then cloned into lentivirus vector pLVX-IRES-ZsGreen1 (Clontech) and pMSCV-puro (Clontech), respectively. .. To construct pMIR-HOXB3 3′UTR plasmid, human HOXB3 3′UTR was amplified by PCR and cloned into pMIR-REPORT vector (Ambion, Dallas, TX, USA).

    Article Title: A novel miR-375-HOXB3-CDCA3/DNMT3B regulatory circuitry contributes to leukemogenesis in acute myeloid leukemia
    Article Snippet: .. To produce plasmids expressing HOXB3 and DNMT3B, human HOXB3 and DNMT3B coding sequences were amplified by PCR and then cloned into lentivirus vector pLVX-IRES-ZsGreen1 (Clontech) and pMSCV-puro (Clontech), respectively. .. To construct pMIR-HOXB3 3′UTR plasmid, human HOXB3 3′UTR was amplified by PCR and cloned into pMIR-REPORT vector (Ambion, Dallas, TX, USA).

    Article Title: Identification of the Intragenomic Promoter Controlling Hepatitis E Virus Subgenomic RNA Transcription
    Article Snippet: .. To construct lentiviral constructs encoding ORF1 of Kernow C1/p6 (GenBank accession number JQ679013 ), the Kernow C1/p6 ORF1 cDNA was amplified by PCR from a plasmid encoding the full-length (FL) infectious HEV clone Kernow C1/p6 (a kind gift from Suzanne Emerson, NIH) and then cloned into pLVX-IRES-zsGreen1 vector using an In-Fusion HD cloning kit (Clontech, Mountain View, CA, USA). .. The GAD mutant of ORF1 inactivating the polymerase was generated by QuikChange (Stratagene) site-directed mutagenesis.

    Construct:

    Article Title: Hepatitis E virus ORF3 is a functional ion channel required for release of infectious particles
    Article Snippet: .. To construct a lentiviral construct that encodes Kernow C1/p6 ORF2 or Kernow C1/p6 ORF3, the Kernow C1/p6 ORF2 or Kernow C1/p6 ORF3 cDNA was amplified by PCR from the HEV Kernow C1/p6 construct (a kind gift from Suzanne Emerson, NIH, Bethesda, MD) and then cloned into pLVX-IRES-zsGreen1 or pLEX-IRES-mCherry vectors using the In-Fusion HD Cloning Kit (Clontech). .. To construct the pLEX-IAV M2-IRES-mCherry vector, cDNA encoding Influenza A virus M2 (A/Puerto Rico/8/34/Mount Sinai/Wi(H1N1)) was synthesized by IDT with gBlock, and then cloned into pLEX-IRES-mCherry vectors using In-Fusion HD Cloning Kit (Clontech).

    Article Title: Identification of the Intragenomic Promoter Controlling Hepatitis E Virus Subgenomic RNA Transcription
    Article Snippet: .. To construct lentiviral constructs encoding ORF1 of Kernow C1/p6 (GenBank accession number JQ679013 ), the Kernow C1/p6 ORF1 cDNA was amplified by PCR from a plasmid encoding the full-length (FL) infectious HEV clone Kernow C1/p6 (a kind gift from Suzanne Emerson, NIH) and then cloned into pLVX-IRES-zsGreen1 vector using an In-Fusion HD cloning kit (Clontech, Mountain View, CA, USA). .. The GAD mutant of ORF1 inactivating the polymerase was generated by QuikChange (Stratagene) site-directed mutagenesis.

    Mouse Assay:

    Article Title: A novel miR-375-HOXB3-CDCA3/DNMT3B regulatory circuitry contributes to leukemogenesis in acute myeloid leukemia
    Article Snippet: .. Engraftment of NOD/SCID-IL2Rγ mice (NSG) THP1 cells were transduced with lentivirus vector pLVX-IRES-ZsGreen1 (Clontech), followed by sorting to obtain GFP-positive cells. .. GFP-positive THP1 cells were transduced with MSCV-miR-375 or MSCV-NC, followed by puromycin selection for 1 week.

    Expressing:

    Article Title: A novel miR-375-HOXB3-CDCA3/DNMT3B regulatory circuitry contributes to leukemogenesis in acute myeloid leukemia
    Article Snippet: .. To produce plasmids expressing HOXB3 and DNMT3B, human HOXB3 and DNMT3B coding sequences were amplified by PCR and then cloned into lentivirus vector pLVX-IRES-ZsGreen1 (Clontech) and pMSCV-puro (Clontech), respectively. .. To construct pMIR-HOXB3 3′UTR plasmid, human HOXB3 3′UTR was amplified by PCR and cloned into pMIR-REPORT vector (Ambion, Dallas, TX, USA).

    Article Title: Selection of an optimal promoter for gene transfer in normal B cells
    Article Snippet: .. B cells activated with CD40L and IL-21 are resistant to transduction To determine the levels of transgene expression in B cells three bicistronic plasmids were used that encode various GFPs under the control of different promoters, namely pGIPZ [cytomegalovirus (CMV) promoter; turbo GFP, which is an improved variant of the green fluorescent protein CopGFP], pLVTHM [elongation factor 1 alpha (EF1α) promoter; GFP) and pLVX-IRES-ZsGreen1 (CMV promoter; ZsGreen1, which is a human codon-optimized variant of ZsGreen) ( ). .. Independent of the vectors used in the present study, a very low level of transgene expression was detected in B cells; expression did not exceed 10%, as assessed with flow cytometry 7 days post-transduction ( ).

    Article Title: A novel miR-375-HOXB3-CDCA3/DNMT3B regulatory circuitry contributes to leukemogenesis in acute myeloid leukemia
    Article Snippet: .. To produce plasmids expressing HOXB3 and DNMT3B, human HOXB3 and DNMT3B coding sequences were amplified by PCR and then cloned into lentivirus vector pLVX-IRES-ZsGreen1 (Clontech) and pMSCV-puro (Clontech), respectively. .. To construct pMIR-HOXB3 3′UTR plasmid, human HOXB3 3′UTR was amplified by PCR and cloned into pMIR-REPORT vector (Ambion, Dallas, TX, USA).

    Polymerase Chain Reaction:

    Article Title: Hepatitis E virus ORF3 is a functional ion channel required for release of infectious particles
    Article Snippet: .. To construct a lentiviral construct that encodes Kernow C1/p6 ORF2 or Kernow C1/p6 ORF3, the Kernow C1/p6 ORF2 or Kernow C1/p6 ORF3 cDNA was amplified by PCR from the HEV Kernow C1/p6 construct (a kind gift from Suzanne Emerson, NIH, Bethesda, MD) and then cloned into pLVX-IRES-zsGreen1 or pLEX-IRES-mCherry vectors using the In-Fusion HD Cloning Kit (Clontech). .. To construct the pLEX-IAV M2-IRES-mCherry vector, cDNA encoding Influenza A virus M2 (A/Puerto Rico/8/34/Mount Sinai/Wi(H1N1)) was synthesized by IDT with gBlock, and then cloned into pLEX-IRES-mCherry vectors using In-Fusion HD Cloning Kit (Clontech).

    Article Title: A novel miR-375-HOXB3-CDCA3/DNMT3B regulatory circuitry contributes to leukemogenesis in acute myeloid leukemia
    Article Snippet: .. To produce plasmids expressing HOXB3 and DNMT3B, human HOXB3 and DNMT3B coding sequences were amplified by PCR and then cloned into lentivirus vector pLVX-IRES-ZsGreen1 (Clontech) and pMSCV-puro (Clontech), respectively. .. To construct pMIR-HOXB3 3′UTR plasmid, human HOXB3 3′UTR was amplified by PCR and cloned into pMIR-REPORT vector (Ambion, Dallas, TX, USA).

    Article Title: A novel miR-375-HOXB3-CDCA3/DNMT3B regulatory circuitry contributes to leukemogenesis in acute myeloid leukemia
    Article Snippet: .. To produce plasmids expressing HOXB3 and DNMT3B, human HOXB3 and DNMT3B coding sequences were amplified by PCR and then cloned into lentivirus vector pLVX-IRES-ZsGreen1 (Clontech) and pMSCV-puro (Clontech), respectively. .. To construct pMIR-HOXB3 3′UTR plasmid, human HOXB3 3′UTR was amplified by PCR and cloned into pMIR-REPORT vector (Ambion, Dallas, TX, USA).

    Article Title: Identification of the Intragenomic Promoter Controlling Hepatitis E Virus Subgenomic RNA Transcription
    Article Snippet: .. To construct lentiviral constructs encoding ORF1 of Kernow C1/p6 (GenBank accession number JQ679013 ), the Kernow C1/p6 ORF1 cDNA was amplified by PCR from a plasmid encoding the full-length (FL) infectious HEV clone Kernow C1/p6 (a kind gift from Suzanne Emerson, NIH) and then cloned into pLVX-IRES-zsGreen1 vector using an In-Fusion HD cloning kit (Clontech, Mountain View, CA, USA). .. The GAD mutant of ORF1 inactivating the polymerase was generated by QuikChange (Stratagene) site-directed mutagenesis.

    Variant Assay:

    Article Title: Selection of an optimal promoter for gene transfer in normal B cells
    Article Snippet: .. B cells activated with CD40L and IL-21 are resistant to transduction To determine the levels of transgene expression in B cells three bicistronic plasmids were used that encode various GFPs under the control of different promoters, namely pGIPZ [cytomegalovirus (CMV) promoter; turbo GFP, which is an improved variant of the green fluorescent protein CopGFP], pLVTHM [elongation factor 1 alpha (EF1α) promoter; GFP) and pLVX-IRES-ZsGreen1 (CMV promoter; ZsGreen1, which is a human codon-optimized variant of ZsGreen) ( ). .. Independent of the vectors used in the present study, a very low level of transgene expression was detected in B cells; expression did not exceed 10%, as assessed with flow cytometry 7 days post-transduction ( ).

    Plasmid Preparation:

    Article Title: A novel miR-375-HOXB3-CDCA3/DNMT3B regulatory circuitry contributes to leukemogenesis in acute myeloid leukemia
    Article Snippet: .. To produce plasmids expressing HOXB3 and DNMT3B, human HOXB3 and DNMT3B coding sequences were amplified by PCR and then cloned into lentivirus vector pLVX-IRES-ZsGreen1 (Clontech) and pMSCV-puro (Clontech), respectively. .. To construct pMIR-HOXB3 3′UTR plasmid, human HOXB3 3′UTR was amplified by PCR and cloned into pMIR-REPORT vector (Ambion, Dallas, TX, USA).

    Article Title: A novel miR-375-HOXB3-CDCA3/DNMT3B regulatory circuitry contributes to leukemogenesis in acute myeloid leukemia
    Article Snippet: .. Engraftment of NOD/SCID-IL2Rγ mice (NSG) THP1 cells were transduced with lentivirus vector pLVX-IRES-ZsGreen1 (Clontech), followed by sorting to obtain GFP-positive cells. .. GFP-positive THP1 cells were transduced with MSCV-miR-375 or MSCV-NC, followed by puromycin selection for 1 week.

    Article Title: A novel miR-375-HOXB3-CDCA3/DNMT3B regulatory circuitry contributes to leukemogenesis in acute myeloid leukemia
    Article Snippet: .. To produce plasmids expressing HOXB3 and DNMT3B, human HOXB3 and DNMT3B coding sequences were amplified by PCR and then cloned into lentivirus vector pLVX-IRES-ZsGreen1 (Clontech) and pMSCV-puro (Clontech), respectively. .. To construct pMIR-HOXB3 3′UTR plasmid, human HOXB3 3′UTR was amplified by PCR and cloned into pMIR-REPORT vector (Ambion, Dallas, TX, USA).

    Article Title: Identification of the Intragenomic Promoter Controlling Hepatitis E Virus Subgenomic RNA Transcription
    Article Snippet: .. To construct lentiviral constructs encoding ORF1 of Kernow C1/p6 (GenBank accession number JQ679013 ), the Kernow C1/p6 ORF1 cDNA was amplified by PCR from a plasmid encoding the full-length (FL) infectious HEV clone Kernow C1/p6 (a kind gift from Suzanne Emerson, NIH) and then cloned into pLVX-IRES-zsGreen1 vector using an In-Fusion HD cloning kit (Clontech, Mountain View, CA, USA). .. The GAD mutant of ORF1 inactivating the polymerase was generated by QuikChange (Stratagene) site-directed mutagenesis.

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    TaKaRa lentivirus vector plvx ires zsgreen1
    The anti-leukemia effects of miR-375 in vivo. About 1х10 7 viable HL-60 cells transduced with MSCV-miR-375 or MSCV-NC were injected subcutaneously into right flank of each nude mouse. a A photograph of xenografted tumors in mice xenografted by HL-60 cells, which were transduced with MSCV-miR-375 or MSCV-NC. b Volumes of all xenografted tumors were measured when the experiment was terminated at 42 days after tumor cell inoculation. c Net weights of all xenografted tumors were measured at the termination of the experiment. d The protein expression of HOXB3 was detected in xenografted tumor lysates from HL-60 cells transduced with MSCV-miR-375 or MSCV-NC. e THP1 cells were transduced with <t>lentivirus</t> vector <t>pLVX-IRES-ZsGreen1</t> and GFP-positive cells were sorted by flow cytometry. The GFP-positive cells were further transduced with MSCV-miR-375 or MSCV-NC and treated with puromycin for 1 week. Then, THP1-GFP-miR-375 or THP1-GFP-NC were xenografted into NSG mice. Peripheral blood cells were extracted from mice when the mice became moribund and GFP-positive cells were analyzed by flow cytometry. * P
    Lentivirus Vector Plvx Ires Zsgreen1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    The anti-leukemia effects of miR-375 in vivo. About 1х10 7 viable HL-60 cells transduced with MSCV-miR-375 or MSCV-NC were injected subcutaneously into right flank of each nude mouse. a A photograph of xenografted tumors in mice xenografted by HL-60 cells, which were transduced with MSCV-miR-375 or MSCV-NC. b Volumes of all xenografted tumors were measured when the experiment was terminated at 42 days after tumor cell inoculation. c Net weights of all xenografted tumors were measured at the termination of the experiment. d The protein expression of HOXB3 was detected in xenografted tumor lysates from HL-60 cells transduced with MSCV-miR-375 or MSCV-NC. e THP1 cells were transduced with lentivirus vector pLVX-IRES-ZsGreen1 and GFP-positive cells were sorted by flow cytometry. The GFP-positive cells were further transduced with MSCV-miR-375 or MSCV-NC and treated with puromycin for 1 week. Then, THP1-GFP-miR-375 or THP1-GFP-NC were xenografted into NSG mice. Peripheral blood cells were extracted from mice when the mice became moribund and GFP-positive cells were analyzed by flow cytometry. * P

    Journal: BMC Cancer

    Article Title: A novel miR-375-HOXB3-CDCA3/DNMT3B regulatory circuitry contributes to leukemogenesis in acute myeloid leukemia

    doi: 10.1186/s12885-018-4097-z

    Figure Lengend Snippet: The anti-leukemia effects of miR-375 in vivo. About 1х10 7 viable HL-60 cells transduced with MSCV-miR-375 or MSCV-NC were injected subcutaneously into right flank of each nude mouse. a A photograph of xenografted tumors in mice xenografted by HL-60 cells, which were transduced with MSCV-miR-375 or MSCV-NC. b Volumes of all xenografted tumors were measured when the experiment was terminated at 42 days after tumor cell inoculation. c Net weights of all xenografted tumors were measured at the termination of the experiment. d The protein expression of HOXB3 was detected in xenografted tumor lysates from HL-60 cells transduced with MSCV-miR-375 or MSCV-NC. e THP1 cells were transduced with lentivirus vector pLVX-IRES-ZsGreen1 and GFP-positive cells were sorted by flow cytometry. The GFP-positive cells were further transduced with MSCV-miR-375 or MSCV-NC and treated with puromycin for 1 week. Then, THP1-GFP-miR-375 or THP1-GFP-NC were xenografted into NSG mice. Peripheral blood cells were extracted from mice when the mice became moribund and GFP-positive cells were analyzed by flow cytometry. * P

    Article Snippet: To produce plasmids expressing HOXB3 and DNMT3B, human HOXB3 and DNMT3B coding sequences were amplified by PCR and then cloned into lentivirus vector pLVX-IRES-ZsGreen1 (Clontech) and pMSCV-puro (Clontech), respectively.

    Techniques: In Vivo, Transduction, Injection, Mouse Assay, Expressing, Plasmid Preparation, Flow Cytometry, Cytometry

    The anti-leukemia effects of miR-375 in vivo. About 1х10 7 viable HL-60 cells transduced with MSCV-miR-375 or MSCV-NC were injected subcutaneously into right flank of each nude mouse. a A photograph of xenografted tumors in mice xenografted by HL-60 cells, which were transduced with MSCV-miR-375 or MSCV-NC. b Volumes of all xenografted tumors were measured when the experiment was terminated at 42 days after tumor cell inoculation. c Net weights of all xenografted tumors were measured at the termination of the experiment. d The protein expression of HOXB3 was detected in xenografted tumor lysates from HL-60 cells transduced with MSCV-miR-375 or MSCV-NC. e THP1 cells were transduced with lentivirus vector pLVX-IRES-ZsGreen1 and GFP-positive cells were sorted by flow cytometry. The GFP-positive cells were further transduced with MSCV-miR-375 or MSCV-NC and treated with puromycin for 1 week. Then, THP1-GFP-miR-375 or THP1-GFP-NC were xenografted into NSG mice. Peripheral blood cells were extracted from mice when the mice became moribund and GFP-positive cells were analyzed by flow cytometry. * P

    Journal: BMC Cancer

    Article Title: A novel miR-375-HOXB3-CDCA3/DNMT3B regulatory circuitry contributes to leukemogenesis in acute myeloid leukemia

    doi: 10.1186/s12885-018-4097-z

    Figure Lengend Snippet: The anti-leukemia effects of miR-375 in vivo. About 1х10 7 viable HL-60 cells transduced with MSCV-miR-375 or MSCV-NC were injected subcutaneously into right flank of each nude mouse. a A photograph of xenografted tumors in mice xenografted by HL-60 cells, which were transduced with MSCV-miR-375 or MSCV-NC. b Volumes of all xenografted tumors were measured when the experiment was terminated at 42 days after tumor cell inoculation. c Net weights of all xenografted tumors were measured at the termination of the experiment. d The protein expression of HOXB3 was detected in xenografted tumor lysates from HL-60 cells transduced with MSCV-miR-375 or MSCV-NC. e THP1 cells were transduced with lentivirus vector pLVX-IRES-ZsGreen1 and GFP-positive cells were sorted by flow cytometry. The GFP-positive cells were further transduced with MSCV-miR-375 or MSCV-NC and treated with puromycin for 1 week. Then, THP1-GFP-miR-375 or THP1-GFP-NC were xenografted into NSG mice. Peripheral blood cells were extracted from mice when the mice became moribund and GFP-positive cells were analyzed by flow cytometry. * P

    Article Snippet: To produce plasmids expressing HOXB3 and DNMT3B, human HOXB3 and DNMT3B coding sequences were amplified by PCR and then cloned into lentivirus vector pLVX-IRES-ZsGreen1 (Clontech) and pMSCV-puro (Clontech), respectively.

    Techniques: In Vivo, Transduction, Injection, Mouse Assay, Expressing, Plasmid Preparation, Flow Cytometry, Cytometry

    Transduction of human CD4 + primary cells with MLV-A-based retroviral VLPs conveying NB-ZsGreen1 or ZsGreen1. (a) Schematic of the methods used to generate CD4 + primary cells transductants. (b) NB-ZsGreen1 or ZsGreen1 VLPs were used to transduce activated

    Journal: Human Gene Therapy

    Article Title: A Mutant Tat Protein Provides Strong Protection from HIV-1 Infection in Human CD4+ T Cells

    doi: 10.1089/hum.2012.176

    Figure Lengend Snippet: Transduction of human CD4 + primary cells with MLV-A-based retroviral VLPs conveying NB-ZsGreen1 or ZsGreen1. (a) Schematic of the methods used to generate CD4 + primary cells transductants. (b) NB-ZsGreen1 or ZsGreen1 VLPs were used to transduce activated

    Article Snippet: Similarly, pGCsamEN-EGFP was made by PCR of an EGFP cassette from pCDN3.1-EGFP, using primers P2 and P3, and pGCsamEN-ZsGreen1 was made by PCR of ZsGreen1 from pLVX-IRES-ZsGreen1 (Clontech, Mountain View, CA), using primers P4 and P5.

    Techniques: Transduction

    HIV replication is inhibited in activated human CD4 + primary cells expressing NB-ZsGreen1. (a) The two CD4 + populations described in were infected with HIV-1 89.6 containing 20 ng of CAp24 and grown for 21 days. Cell-free supernatant was

    Journal: Human Gene Therapy

    Article Title: A Mutant Tat Protein Provides Strong Protection from HIV-1 Infection in Human CD4+ T Cells

    doi: 10.1089/hum.2012.176

    Figure Lengend Snippet: HIV replication is inhibited in activated human CD4 + primary cells expressing NB-ZsGreen1. (a) The two CD4 + populations described in were infected with HIV-1 89.6 containing 20 ng of CAp24 and grown for 21 days. Cell-free supernatant was

    Article Snippet: Similarly, pGCsamEN-EGFP was made by PCR of an EGFP cassette from pCDN3.1-EGFP, using primers P2 and P3, and pGCsamEN-ZsGreen1 was made by PCR of ZsGreen1 from pLVX-IRES-ZsGreen1 (Clontech, Mountain View, CA), using primers P4 and P5.

    Techniques: Expressing, Infection

    Neural stem cells (NSC) produce and deliver BiTE LLON protein to tumors in vivo . BiTE transduced NSCs were sorted for the expression of A) ZsGreen1 (sample histogram) protein and evaluated for the production of BiTE proteins using immunocytochemistry by B) NSC LLON and C) NSC LLOFF cells immobilized on cover glass (cells-DAPI-nucleus(I), NSC-ZsGreen1(II), BiTE protein-α-His-Tag (III), and (IV) merged together, scale bar 50μm), and D) western blotting using anti-His Tag and anti-GAPDH antibodies. E) NSC LLON but not NSC LLOFF produced protein binding to IL13Rα2 protein immobilized on ELISA plates. F) Production dynamics of BiTE LLON by NSC LLON after 24 and 48 hours in culture (T-test, n=3, **p

    Journal: bioRxiv

    Article Title: Neural Stem Cells Secreting Bispecific T Cell Engager to Induce Selective Anti-Glioma Activity

    doi: 10.1101/2020.07.21.188441

    Figure Lengend Snippet: Neural stem cells (NSC) produce and deliver BiTE LLON protein to tumors in vivo . BiTE transduced NSCs were sorted for the expression of A) ZsGreen1 (sample histogram) protein and evaluated for the production of BiTE proteins using immunocytochemistry by B) NSC LLON and C) NSC LLOFF cells immobilized on cover glass (cells-DAPI-nucleus(I), NSC-ZsGreen1(II), BiTE protein-α-His-Tag (III), and (IV) merged together, scale bar 50μm), and D) western blotting using anti-His Tag and anti-GAPDH antibodies. E) NSC LLON but not NSC LLOFF produced protein binding to IL13Rα2 protein immobilized on ELISA plates. F) Production dynamics of BiTE LLON by NSC LLON after 24 and 48 hours in culture (T-test, n=3, **p

    Article Snippet: Generation of 293T and neural stem cells secreting BiTE protein The pLVX-IRES-ZsGreen1 plasmid (Takara Bio USA, Inc., Mountain View, CA) was used to generate construct encoding BiTE cDNA.

    Techniques: In Vivo, Expressing, Immunocytochemistry, Western Blot, Produced, Protein Binding, Enzyme-linked Immunosorbent Assay, T-Test