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TaKaRa plasmid cmv pegfp c1
Expression of EGFP protein in cells transiently transfected with EGFP under the control of PABP3 promoter sequences. PABP3 genomic sequences extending from nt –498 to +30 were amplified by PCR, as described in Materials and Methods, and then inserted into a promoter-less EGFP vector <t>(pEGFP-1).</t> These constructs were transiently transfected into HeLa (black boxes) or NTERA-2 (grey boxes) cells. The numbers of cells counted were identical in the two transfection assays. EGFP fluorescence driven by each construct was normalised to that obtained with the <t>CMV</t> <t>promoter/pEGFP-C1</t> vector (fluorescence of 100%). Results are the means of triplicate determinations obtained in two independent experiments. CMV corresponds to the control construct of the CMV promoter upstream from EGFP. PC, P1, P2 and P3 refer to the different PABP3 promoter constructs upstream from the EGFP reporter gene as indicated on the left.
Plasmid Cmv Pegfp C1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Human testis expresses a specific poly(A)-binding protein"

Article Title: Human testis expresses a specific poly(A)-binding protein

Journal: Nucleic Acids Research

doi:

Expression of EGFP protein in cells transiently transfected with EGFP under the control of PABP3 promoter sequences. PABP3 genomic sequences extending from nt –498 to +30 were amplified by PCR, as described in Materials and Methods, and then inserted into a promoter-less EGFP vector (pEGFP-1). These constructs were transiently transfected into HeLa (black boxes) or NTERA-2 (grey boxes) cells. The numbers of cells counted were identical in the two transfection assays. EGFP fluorescence driven by each construct was normalised to that obtained with the CMV promoter/pEGFP-C1 vector (fluorescence of 100%). Results are the means of triplicate determinations obtained in two independent experiments. CMV corresponds to the control construct of the CMV promoter upstream from EGFP. PC, P1, P2 and P3 refer to the different PABP3 promoter constructs upstream from the EGFP reporter gene as indicated on the left.
Figure Legend Snippet: Expression of EGFP protein in cells transiently transfected with EGFP under the control of PABP3 promoter sequences. PABP3 genomic sequences extending from nt –498 to +30 were amplified by PCR, as described in Materials and Methods, and then inserted into a promoter-less EGFP vector (pEGFP-1). These constructs were transiently transfected into HeLa (black boxes) or NTERA-2 (grey boxes) cells. The numbers of cells counted were identical in the two transfection assays. EGFP fluorescence driven by each construct was normalised to that obtained with the CMV promoter/pEGFP-C1 vector (fluorescence of 100%). Results are the means of triplicate determinations obtained in two independent experiments. CMV corresponds to the control construct of the CMV promoter upstream from EGFP. PC, P1, P2 and P3 refer to the different PABP3 promoter constructs upstream from the EGFP reporter gene as indicated on the left.

Techniques Used: Expressing, Transfection, Genomic Sequencing, Amplification, Polymerase Chain Reaction, Plasmid Preparation, Construct, Fluorescence

Related Articles

Transfection:

Article Title: Human testis expresses a specific poly(A)-binding protein
Article Snippet: .. Plasmid CMV/pEGFP-C1 (Clontech), which contains the human cytomegalovirus (CMV) promoter, was used as a positive control in transfection experiments. .. The human NTERA-2/D1 cell line (NTERA-2/clone D1, a human pluripotent embryonic carcinoma cell line derived from a lung metastasis of a testicular teratocarcinoma) was obtained from the American Type Culture Collection (no. CRL 1973; ATCC, Biovalley, France).

Plasmid Preparation:

Article Title: Human testis expresses a specific poly(A)-binding protein
Article Snippet: .. Plasmid CMV/pEGFP-C1 (Clontech), which contains the human cytomegalovirus (CMV) promoter, was used as a positive control in transfection experiments. .. The human NTERA-2/D1 cell line (NTERA-2/clone D1, a human pluripotent embryonic carcinoma cell line derived from a lung metastasis of a testicular teratocarcinoma) was obtained from the American Type Culture Collection (no. CRL 1973; ATCC, Biovalley, France).

Positive Control:

Article Title: Human testis expresses a specific poly(A)-binding protein
Article Snippet: .. Plasmid CMV/pEGFP-C1 (Clontech), which contains the human cytomegalovirus (CMV) promoter, was used as a positive control in transfection experiments. .. The human NTERA-2/D1 cell line (NTERA-2/clone D1, a human pluripotent embryonic carcinoma cell line derived from a lung metastasis of a testicular teratocarcinoma) was obtained from the American Type Culture Collection (no. CRL 1973; ATCC, Biovalley, France).

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    TaKaRa plasmid cmv pegfp c1
    Expression of EGFP protein in cells transiently transfected with EGFP under the control of PABP3 promoter sequences. PABP3 genomic sequences extending from nt –498 to +30 were amplified by PCR, as described in Materials and Methods, and then inserted into a promoter-less EGFP vector <t>(pEGFP-1).</t> These constructs were transiently transfected into HeLa (black boxes) or NTERA-2 (grey boxes) cells. The numbers of cells counted were identical in the two transfection assays. EGFP fluorescence driven by each construct was normalised to that obtained with the <t>CMV</t> <t>promoter/pEGFP-C1</t> vector (fluorescence of 100%). Results are the means of triplicate determinations obtained in two independent experiments. CMV corresponds to the control construct of the CMV promoter upstream from EGFP. PC, P1, P2 and P3 refer to the different PABP3 promoter constructs upstream from the EGFP reporter gene as indicated on the left.
    Plasmid Cmv Pegfp C1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/plasmid cmv pegfp c1/product/TaKaRa
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    plasmid cmv pegfp c1 - by Bioz Stars, 2020-08
    85/100 stars
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    85
    TaKaRa cmv pegfp c1
    Expression of EGFP protein in cells transiently transfected with EGFP under the control of PABP3 promoter sequences. PABP3 genomic sequences extending from nt –498 to +30 were amplified by PCR, as described in Materials and Methods, and then inserted into a promoter-less EGFP vector <t>(pEGFP-1).</t> These constructs were transiently transfected into HeLa (black boxes) or NTERA-2 (grey boxes) cells. The numbers of cells counted were identical in the two transfection assays. EGFP fluorescence driven by each construct was normalised to that obtained with the <t>CMV</t> <t>promoter/pEGFP-C1</t> vector (fluorescence of 100%). Results are the means of triplicate determinations obtained in two independent experiments. CMV corresponds to the control construct of the CMV promoter upstream from EGFP. PC, P1, P2 and P3 refer to the different PABP3 promoter constructs upstream from the EGFP reporter gene as indicated on the left.
    Cmv Pegfp C1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cmv pegfp c1/product/TaKaRa
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cmv pegfp c1 - by Bioz Stars, 2020-08
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    90
    TaKaRa pzdonor egfp
    AAVS1 locus-specific integration of different size donor DNAs . ( a ) ZFN-dependent integration of donor DNA. K562 cells were coelectroporated with pEGFP (reporter for transfection efficiency) and <t>pZDonor</t> with or without AAVS1 ZFN mRNA. (Left): Brightfield and fluorescence images of transfected cells. Bar = 100 µm (Right): PCR spanning the integration junction (top) and RFLP assay (bottom) performed on genomic DNA from cells 4 days after treatment with pZdonor only; or 4 (Day 4) and 16 days (Day 16) after treatment with pZdonor and AAVS1 ZFN mRNA provided evidence of site-specific integration of 50-bp donor DNA. Control PCR amplified a 900-bp region of the AAVS1 locus. ( b ) Accuracy of Enhanced Sharkey AAVS1 ZFN-mediated integration of pZdonor <t>EGFP.</t> Left: PCR amplification of the left and right integration junctions performed on genomic DNA of K562 cells coelectroporated with pZDonor EGFP and Enhanced Sharkey ZFN with or without G418 selection. Right: DNA sequence chromatogram of left (top) and right (bottom) junctional PCR amplicons. Vector sequences are underlined in blue; Enhanced Sharkey AAVS1 ZFN recognition half-sites are underlined in red. Control PCR amplified a 900-bp region of the AAVS1 locus. WT K562 denotes untransfected control K562 cells. ( c ) Accuracy of Enhanced Sharkey AAVS1 ZFN-mediated integration of pZDonor Hybrid FVIII. Left: PCR amplification of the left and right integration junctions performed on genomic DNA of K562 cells electroporated with pZDonor Hybrid FVIII only or coelectroporated with Enhanced Sharkey ZFN followed by G418 selection. Right: DNA sequence chromatogram of left (top) and right (bottom) junctional PCR amplicons. Vector sequences are underlined in blue; Enhanced Sharkey AAVS1 ZFN recognition half-sites are underlined in red. Control PCR amplified a 900-bp region of the AAVS1 locus. WT K562 denotes untransfected control K562 cells. White vertical lines in the gel images demarcate lanes that were merged for clarity.
    Pzdonor Egfp, supplied by TaKaRa, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    TaKaRa pegfp c1
    AAVS1 locus-specific integration of different size donor DNAs . ( a ) ZFN-dependent integration of donor DNA. K562 cells were coelectroporated with pEGFP (reporter for transfection efficiency) and <t>pZDonor</t> with or without AAVS1 ZFN mRNA. (Left): Brightfield and fluorescence images of transfected cells. Bar = 100 µm (Right): PCR spanning the integration junction (top) and RFLP assay (bottom) performed on genomic DNA from cells 4 days after treatment with pZdonor only; or 4 (Day 4) and 16 days (Day 16) after treatment with pZdonor and AAVS1 ZFN mRNA provided evidence of site-specific integration of 50-bp donor DNA. Control PCR amplified a 900-bp region of the AAVS1 locus. ( b ) Accuracy of Enhanced Sharkey AAVS1 ZFN-mediated integration of pZdonor <t>EGFP.</t> Left: PCR amplification of the left and right integration junctions performed on genomic DNA of K562 cells coelectroporated with pZDonor EGFP and Enhanced Sharkey ZFN with or without G418 selection. Right: DNA sequence chromatogram of left (top) and right (bottom) junctional PCR amplicons. Vector sequences are underlined in blue; Enhanced Sharkey AAVS1 ZFN recognition half-sites are underlined in red. Control PCR amplified a 900-bp region of the AAVS1 locus. WT K562 denotes untransfected control K562 cells. ( c ) Accuracy of Enhanced Sharkey AAVS1 ZFN-mediated integration of pZDonor Hybrid FVIII. Left: PCR amplification of the left and right integration junctions performed on genomic DNA of K562 cells electroporated with pZDonor Hybrid FVIII only or coelectroporated with Enhanced Sharkey ZFN followed by G418 selection. Right: DNA sequence chromatogram of left (top) and right (bottom) junctional PCR amplicons. Vector sequences are underlined in blue; Enhanced Sharkey AAVS1 ZFN recognition half-sites are underlined in red. Control PCR amplified a 900-bp region of the AAVS1 locus. WT K562 denotes untransfected control K562 cells. White vertical lines in the gel images demarcate lanes that were merged for clarity.
    Pegfp C1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 94/100, based on 1379 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Expression of EGFP protein in cells transiently transfected with EGFP under the control of PABP3 promoter sequences. PABP3 genomic sequences extending from nt –498 to +30 were amplified by PCR, as described in Materials and Methods, and then inserted into a promoter-less EGFP vector (pEGFP-1). These constructs were transiently transfected into HeLa (black boxes) or NTERA-2 (grey boxes) cells. The numbers of cells counted were identical in the two transfection assays. EGFP fluorescence driven by each construct was normalised to that obtained with the CMV promoter/pEGFP-C1 vector (fluorescence of 100%). Results are the means of triplicate determinations obtained in two independent experiments. CMV corresponds to the control construct of the CMV promoter upstream from EGFP. PC, P1, P2 and P3 refer to the different PABP3 promoter constructs upstream from the EGFP reporter gene as indicated on the left.

    Journal: Nucleic Acids Research

    Article Title: Human testis expresses a specific poly(A)-binding protein

    doi:

    Figure Lengend Snippet: Expression of EGFP protein in cells transiently transfected with EGFP under the control of PABP3 promoter sequences. PABP3 genomic sequences extending from nt –498 to +30 were amplified by PCR, as described in Materials and Methods, and then inserted into a promoter-less EGFP vector (pEGFP-1). These constructs were transiently transfected into HeLa (black boxes) or NTERA-2 (grey boxes) cells. The numbers of cells counted were identical in the two transfection assays. EGFP fluorescence driven by each construct was normalised to that obtained with the CMV promoter/pEGFP-C1 vector (fluorescence of 100%). Results are the means of triplicate determinations obtained in two independent experiments. CMV corresponds to the control construct of the CMV promoter upstream from EGFP. PC, P1, P2 and P3 refer to the different PABP3 promoter constructs upstream from the EGFP reporter gene as indicated on the left.

    Article Snippet: Plasmid CMV/pEGFP-C1 (Clontech), which contains the human cytomegalovirus (CMV) promoter, was used as a positive control in transfection experiments.

    Techniques: Expressing, Transfection, Genomic Sequencing, Amplification, Polymerase Chain Reaction, Plasmid Preparation, Construct, Fluorescence

    Expression of EGFP protein in cells transiently transfected with EGFP under the control of PABP3 promoter sequences. PABP3 genomic sequences extending from nt –498 to +30 were amplified by PCR, as described in Materials and Methods, and then inserted into a promoter-less EGFP vector (pEGFP-1). These constructs were transiently transfected into HeLa (black boxes) or NTERA-2 (grey boxes) cells. The numbers of cells counted were identical in the two transfection assays. EGFP fluorescence driven by each construct was normalised to that obtained with the CMV promoter/pEGFP-C1 vector (fluorescence of 100%). Results are the means of triplicate determinations obtained in two independent experiments. CMV corresponds to the control construct of the CMV promoter upstream from EGFP. PC, P1, P2 and P3 refer to the different PABP3 promoter constructs upstream from the EGFP reporter gene as indicated on the left.

    Journal: Nucleic Acids Research

    Article Title: Human testis expresses a specific poly(A)-binding protein

    doi:

    Figure Lengend Snippet: Expression of EGFP protein in cells transiently transfected with EGFP under the control of PABP3 promoter sequences. PABP3 genomic sequences extending from nt –498 to +30 were amplified by PCR, as described in Materials and Methods, and then inserted into a promoter-less EGFP vector (pEGFP-1). These constructs were transiently transfected into HeLa (black boxes) or NTERA-2 (grey boxes) cells. The numbers of cells counted were identical in the two transfection assays. EGFP fluorescence driven by each construct was normalised to that obtained with the CMV promoter/pEGFP-C1 vector (fluorescence of 100%). Results are the means of triplicate determinations obtained in two independent experiments. CMV corresponds to the control construct of the CMV promoter upstream from EGFP. PC, P1, P2 and P3 refer to the different PABP3 promoter constructs upstream from the EGFP reporter gene as indicated on the left.

    Article Snippet: They were then transfected with 400 ng of either CMV/pEGFP-C1 (Clontech) as a positive control or the PABP3/pEGFP-1 constructs with 4 µl of LipofectAMINE and 1 µl of PLUS reagent (Life Technologies) in 200 µl of serum-free medium.

    Techniques: Expressing, Transfection, Genomic Sequencing, Amplification, Polymerase Chain Reaction, Plasmid Preparation, Construct, Fluorescence

    AAVS1 locus-specific integration of different size donor DNAs . ( a ) ZFN-dependent integration of donor DNA. K562 cells were coelectroporated with pEGFP (reporter for transfection efficiency) and pZDonor with or without AAVS1 ZFN mRNA. (Left): Brightfield and fluorescence images of transfected cells. Bar = 100 µm (Right): PCR spanning the integration junction (top) and RFLP assay (bottom) performed on genomic DNA from cells 4 days after treatment with pZdonor only; or 4 (Day 4) and 16 days (Day 16) after treatment with pZdonor and AAVS1 ZFN mRNA provided evidence of site-specific integration of 50-bp donor DNA. Control PCR amplified a 900-bp region of the AAVS1 locus. ( b ) Accuracy of Enhanced Sharkey AAVS1 ZFN-mediated integration of pZdonor EGFP. Left: PCR amplification of the left and right integration junctions performed on genomic DNA of K562 cells coelectroporated with pZDonor EGFP and Enhanced Sharkey ZFN with or without G418 selection. Right: DNA sequence chromatogram of left (top) and right (bottom) junctional PCR amplicons. Vector sequences are underlined in blue; Enhanced Sharkey AAVS1 ZFN recognition half-sites are underlined in red. Control PCR amplified a 900-bp region of the AAVS1 locus. WT K562 denotes untransfected control K562 cells. ( c ) Accuracy of Enhanced Sharkey AAVS1 ZFN-mediated integration of pZDonor Hybrid FVIII. Left: PCR amplification of the left and right integration junctions performed on genomic DNA of K562 cells electroporated with pZDonor Hybrid FVIII only or coelectroporated with Enhanced Sharkey ZFN followed by G418 selection. Right: DNA sequence chromatogram of left (top) and right (bottom) junctional PCR amplicons. Vector sequences are underlined in blue; Enhanced Sharkey AAVS1 ZFN recognition half-sites are underlined in red. Control PCR amplified a 900-bp region of the AAVS1 locus. WT K562 denotes untransfected control K562 cells. White vertical lines in the gel images demarcate lanes that were merged for clarity.

    Journal: Molecular Therapy

    Article Title: Multidimensional Genome-wide Analyses Show Accurate FVIII Integration by ZFN in Primary Human Cells

    doi: 10.1038/mt.2015.223

    Figure Lengend Snippet: AAVS1 locus-specific integration of different size donor DNAs . ( a ) ZFN-dependent integration of donor DNA. K562 cells were coelectroporated with pEGFP (reporter for transfection efficiency) and pZDonor with or without AAVS1 ZFN mRNA. (Left): Brightfield and fluorescence images of transfected cells. Bar = 100 µm (Right): PCR spanning the integration junction (top) and RFLP assay (bottom) performed on genomic DNA from cells 4 days after treatment with pZdonor only; or 4 (Day 4) and 16 days (Day 16) after treatment with pZdonor and AAVS1 ZFN mRNA provided evidence of site-specific integration of 50-bp donor DNA. Control PCR amplified a 900-bp region of the AAVS1 locus. ( b ) Accuracy of Enhanced Sharkey AAVS1 ZFN-mediated integration of pZdonor EGFP. Left: PCR amplification of the left and right integration junctions performed on genomic DNA of K562 cells coelectroporated with pZDonor EGFP and Enhanced Sharkey ZFN with or without G418 selection. Right: DNA sequence chromatogram of left (top) and right (bottom) junctional PCR amplicons. Vector sequences are underlined in blue; Enhanced Sharkey AAVS1 ZFN recognition half-sites are underlined in red. Control PCR amplified a 900-bp region of the AAVS1 locus. WT K562 denotes untransfected control K562 cells. ( c ) Accuracy of Enhanced Sharkey AAVS1 ZFN-mediated integration of pZDonor Hybrid FVIII. Left: PCR amplification of the left and right integration junctions performed on genomic DNA of K562 cells electroporated with pZDonor Hybrid FVIII only or coelectroporated with Enhanced Sharkey ZFN followed by G418 selection. Right: DNA sequence chromatogram of left (top) and right (bottom) junctional PCR amplicons. Vector sequences are underlined in blue; Enhanced Sharkey AAVS1 ZFN recognition half-sites are underlined in red. Control PCR amplified a 900-bp region of the AAVS1 locus. WT K562 denotes untransfected control K562 cells. White vertical lines in the gel images demarcate lanes that were merged for clarity.

    Article Snippet: Three plasmids having a neomycin resistance marker were used to integrate donor DNAs of increasing sizes into intron 1 of PPP1R12C ( Supplementary Figure S1b ): pZDonor (contains 1.5-kb homology to the AAVS1 locus bisected by a 50-bp multiple cloning site; Sigma-Aldrich) pZDonor EGFP (3.75-kb CMV-promoter-GFP excised from pEGFP-C1 (Clontech, Mountain View, CA) cloned in pZDonor) pZDonor Hybrid FVIII (9.1-kb donor encoding human ferritin light chain promoter-hybrid FVIII cDNA cloned in pZDonor; described below).

    Techniques: Transfection, Fluorescence, Polymerase Chain Reaction, RFLP Assay, Amplification, Selection, Sequencing, Plasmid Preparation