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pgl3-basic luciferase reporter vectors  (Promega)

 
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    Structured Review

    Promega pgl3-basic luciferase reporter vectors
    Pgl3 Basic Luciferase Reporter Vectors, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pgl3-basic+luciferase+reporter-vector/pgl3+basic/pmc12261028-74-13-17
    Average 90 stars, based on 1 article reviews
    pgl3-basic luciferase reporter vectors - by Bioz Stars, 2026-10
    90/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Evidence for NR2F2 /COUP-TFII involvement in human testis development
    Article Snippet: The Lhb promoter sequences from the rat (-207/+ 5) , and the equivalent region in the human (− 207/ + 5) were cloned into pGL3-basic Luciferase reporter-vector (E1751, Promega) using the forward primer containing XhoI restriction site: 5′-CGGGCTCGAGTTCCCAATGTCAGTTAAGC-3′ and the reverse primer containing SacI restriction site: 5′-GGTACCGAGCTCTTCCCAATGTCAGTTAAGC-3′ for the rat sequence, and the forward primer containing XhoI restriction site: 5′-CGGGCTCGAGGTCTCTGCCTCACCTCT-3′ and the reverse primer containing SacI: 5′-GGTACCGAGCTCGTCTCTGCCTCACCTCT-3′ for the human sequence.

    Article Title: Evidence for NR2F2/COUP-TFII involvement in human testis development.
    Article Snippet: The Lhb promoter sequences from the rat (-207/+ 5)33, and the equivalent region in the human (− 207/ + 5) were cloned into pGL3-basic Luciferase reporter-vector (E1751, Promega) using the forward primer containing XhoI restriction site: 5′-CGG GCT CGA GTT CCC AAT GTC AGT TAAGC-3′ and the reverse primer containing SacI restriction site: 5′-GGT ACC GAG CTC TTC CCA ATG TCA GTT AAG C-3′ for the rat sequence, and the forward primer containing XhoI restriction site: 5′-CGG GCT CGA GGT CTC TGC CTC ACC TCT-3′ and the reverse primer containing SacI: 5′-GGT ACC GAG CTC GTC TCT GCC TCA CCTCT-3′ for the human sequence.

    Luciferase:

    Article Title: Evidence for NR2F2 /COUP-TFII involvement in human testis development
    Article Snippet: The Lhb promoter sequences from the rat (-207/+ 5) , and the equivalent region in the human (− 207/ + 5) were cloned into pGL3-basic Luciferase reporter-vector (E1751, Promega) using the forward primer containing XhoI restriction site: 5′-CGGGCTCGAGTTCCCAATGTCAGTTAAGC-3′ and the reverse primer containing SacI restriction site: 5′-GGTACCGAGCTCTTCCCAATGTCAGTTAAGC-3′ for the rat sequence, and the forward primer containing XhoI restriction site: 5′-CGGGCTCGAGGTCTCTGCCTCACCTCT-3′ and the reverse primer containing SacI: 5′-GGTACCGAGCTCGTCTCTGCCTCACCTCT-3′ for the human sequence.

    Article Title: Evidence for NR2F2/COUP-TFII involvement in human testis development.
    Article Snippet: The Lhb promoter sequences from the rat (-207/+ 5)33, and the equivalent region in the human (− 207/ + 5) were cloned into pGL3-basic Luciferase reporter-vector (E1751, Promega) using the forward primer containing XhoI restriction site: 5′-CGG GCT CGA GTT CCC AAT GTC AGT TAAGC-3′ and the reverse primer containing SacI restriction site: 5′-GGT ACC GAG CTC TTC CCA ATG TCA GTT AAG C-3′ for the rat sequence, and the forward primer containing XhoI restriction site: 5′-CGG GCT CGA GGT CTC TGC CTC ACC TCT-3′ and the reverse primer containing SacI: 5′-GGT ACC GAG CTC GTC TCT GCC TCA CCTCT-3′ for the human sequence.

    Sequencing:

    Article Title: Evidence for NR2F2 /COUP-TFII involvement in human testis development
    Article Snippet: The Lhb promoter sequences from the rat (-207/+ 5) , and the equivalent region in the human (− 207/ + 5) were cloned into pGL3-basic Luciferase reporter-vector (E1751, Promega) using the forward primer containing XhoI restriction site: 5′-CGGGCTCGAGTTCCCAATGTCAGTTAAGC-3′ and the reverse primer containing SacI restriction site: 5′-GGTACCGAGCTCTTCCCAATGTCAGTTAAGC-3′ for the rat sequence, and the forward primer containing XhoI restriction site: 5′-CGGGCTCGAGGTCTCTGCCTCACCTCT-3′ and the reverse primer containing SacI: 5′-GGTACCGAGCTCGTCTCTGCCTCACCTCT-3′ for the human sequence.

    Article Title: Evidence for NR2F2/COUP-TFII involvement in human testis development.
    Article Snippet: The Lhb promoter sequences from the rat (-207/+ 5)33, and the equivalent region in the human (− 207/ + 5) were cloned into pGL3-basic Luciferase reporter-vector (E1751, Promega) using the forward primer containing XhoI restriction site: 5′-CGG GCT CGA GTT CCC AAT GTC AGT TAAGC-3′ and the reverse primer containing SacI restriction site: 5′-GGT ACC GAG CTC TTC CCA ATG TCA GTT AAG C-3′ for the rat sequence, and the forward primer containing XhoI restriction site: 5′-CGG GCT CGA GGT CTC TGC CTC ACC TCT-3′ and the reverse primer containing SacI: 5′-GGT ACC GAG CTC GTC TCT GCC TCA CCTCT-3′ for the human sequence.

    Countercurrent Chromatography:

    Article Title: Evidence for NR2F2 /COUP-TFII involvement in human testis development
    Article Snippet: The Lhb promoter sequences from the rat (-207/+ 5) , and the equivalent region in the human (− 207/ + 5) were cloned into pGL3-basic Luciferase reporter-vector (E1751, Promega) using the forward primer containing XhoI restriction site: 5′-CGGGCTCGAGTTCCCAATGTCAGTTAAGC-3′ and the reverse primer containing SacI restriction site: 5′-GGTACCGAGCTCTTCCCAATGTCAGTTAAGC-3′ for the rat sequence, and the forward primer containing XhoI restriction site: 5′-CGGGCTCGAGGTCTCTGCCTCACCTCT-3′ and the reverse primer containing SacI: 5′-GGTACCGAGCTCGTCTCTGCCTCACCTCT-3′ for the human sequence.

    Article Title: Evidence for NR2F2/COUP-TFII involvement in human testis development.
    Article Snippet: The Lhb promoter sequences from the rat (-207/+ 5)33, and the equivalent region in the human (− 207/ + 5) were cloned into pGL3-basic Luciferase reporter-vector (E1751, Promega) using the forward primer containing XhoI restriction site: 5′-CGG GCT CGA GTT CCC AAT GTC AGT TAAGC-3′ and the reverse primer containing SacI restriction site: 5′-GGT ACC GAG CTC TTC CCA ATG TCA GTT AAG C-3′ for the rat sequence, and the forward primer containing XhoI restriction site: 5′-CGG GCT CGA GGT CTC TGC CTC ACC TCT-3′ and the reverse primer containing SacI: 5′-GGT ACC GAG CTC GTC TCT GCC TCA CCTCT-3′ for the human sequence.



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    MAD2 and BUB1B are transcriptionally activated by ERα. (A) Schematic representations showing the estrogen response elements (EREs) in the promoter regions of MAD2 and BUB1B . (B) E2 (17β-estradiol) treatment up-regulates MAD2 and BUB1B promoter activity in MCF-7 cells. The top panel shows the map of MAD2 and BUB1B promoter regions used for <t>luciferase</t> reporter assay. pSS-MAD2 construct contains an 1128-bp region (−1098 to +30 nt) of the MAD2 gene and the pSS-BUB1B construct contains a 1546-bp region (−1454 to +92 nt) of the BUB1B gene, cloned into <t>pGL3</t> basic vector. MCF-7 cells were transiently transfected with pSS-MAD2 (250 ng) and pSS-BUB1B (500 ng), followed by E2 treatment, and proceeded for luciferase reporter assay. **p < 0.005; (two-tailed unpaired Student’s t-test) (C) Ectopic ERα up-regulates MAD2 and BUB1B promoter activity in MDA-MB-231 cells. MDA-MB-231 cells were co-transfected with pSS-MAD2 (250 ng) and pSS-BUB1B (500 ng) along with 1.5 µg of ectopic ERα (pCMV3-N-FLAG-ESR1), followed by E2 treatment and proceeded for luciferase reporter assay and plotted as discussed in (B). *p < 0.05; **p < 0.005; ***p < 0.0005 (two-tailed unpaired Student’s t-test). (D) Deletion of ERE results in a significant decrease in luciferase activity. Map of the two deletion constructs, ΔERE pSS-MAD2, and ΔERE pSS-BUB1B were shown in the top panel. MCF-7 cells were transfected with either ΔERE pSS-MAD2 (250 ng) or ΔERE pSS-BUB1B (500 ng), followed by E2 treatment. Data were plotted as discussed in (B). **p < 0.005; ns, not significant (two-tailed unpaired Student’s t-test). (E) ERα physically recruits to MAD2 and BUB1B promoters. The total cell extract of MCF-7 was prepared and Chromatin immunoprecipitation was done using antibodies specific for ERα and normal IgG as control. Precipitated chromatin was estimated by qRT-PCR. The results are expressed as a percent of input. *p < 0.05; **p < 0.005; (two-tailed unpaired Student’s t-test). (F) Representative scatter plot depicting the Pearson’s correlation analysis between the gene expression values (TPM) of MAD2 and BUB1B , extracted from TCGA BRCA cohort using UCSC Xena database. The Pearson correlation coefficient is reported as the ‘r’ value. ** *p < 0.0005; (two-tailed unpaired Student’s t-test). All the experiments (B-E) were performed in triplicates and error bars were represented as means ± s.d.
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    Image Search Results


    MAD2 and BUB1B are transcriptionally activated by ERα. (A) Schematic representations showing the estrogen response elements (EREs) in the promoter regions of MAD2 and BUB1B . (B) E2 (17β-estradiol) treatment up-regulates MAD2 and BUB1B promoter activity in MCF-7 cells. The top panel shows the map of MAD2 and BUB1B promoter regions used for luciferase reporter assay. pSS-MAD2 construct contains an 1128-bp region (−1098 to +30 nt) of the MAD2 gene and the pSS-BUB1B construct contains a 1546-bp region (−1454 to +92 nt) of the BUB1B gene, cloned into pGL3 basic vector. MCF-7 cells were transiently transfected with pSS-MAD2 (250 ng) and pSS-BUB1B (500 ng), followed by E2 treatment, and proceeded for luciferase reporter assay. **p < 0.005; (two-tailed unpaired Student’s t-test) (C) Ectopic ERα up-regulates MAD2 and BUB1B promoter activity in MDA-MB-231 cells. MDA-MB-231 cells were co-transfected with pSS-MAD2 (250 ng) and pSS-BUB1B (500 ng) along with 1.5 µg of ectopic ERα (pCMV3-N-FLAG-ESR1), followed by E2 treatment and proceeded for luciferase reporter assay and plotted as discussed in (B). *p < 0.05; **p < 0.005; ***p < 0.0005 (two-tailed unpaired Student’s t-test). (D) Deletion of ERE results in a significant decrease in luciferase activity. Map of the two deletion constructs, ΔERE pSS-MAD2, and ΔERE pSS-BUB1B were shown in the top panel. MCF-7 cells were transfected with either ΔERE pSS-MAD2 (250 ng) or ΔERE pSS-BUB1B (500 ng), followed by E2 treatment. Data were plotted as discussed in (B). **p < 0.005; ns, not significant (two-tailed unpaired Student’s t-test). (E) ERα physically recruits to MAD2 and BUB1B promoters. The total cell extract of MCF-7 was prepared and Chromatin immunoprecipitation was done using antibodies specific for ERα and normal IgG as control. Precipitated chromatin was estimated by qRT-PCR. The results are expressed as a percent of input. *p < 0.05; **p < 0.005; (two-tailed unpaired Student’s t-test). (F) Representative scatter plot depicting the Pearson’s correlation analysis between the gene expression values (TPM) of MAD2 and BUB1B , extracted from TCGA BRCA cohort using UCSC Xena database. The Pearson correlation coefficient is reported as the ‘r’ value. ** *p < 0.0005; (two-tailed unpaired Student’s t-test). All the experiments (B-E) were performed in triplicates and error bars were represented as means ± s.d.

    Journal: bioRxiv

    Article Title: Estrogen receptor alpha (ERα) driven trans-regulation of mitotic checkpoint complex (MCC) components affects the clinical outcome of breast cancer

    doi: 10.1101/2024.08.31.610592

    Figure Lengend Snippet: MAD2 and BUB1B are transcriptionally activated by ERα. (A) Schematic representations showing the estrogen response elements (EREs) in the promoter regions of MAD2 and BUB1B . (B) E2 (17β-estradiol) treatment up-regulates MAD2 and BUB1B promoter activity in MCF-7 cells. The top panel shows the map of MAD2 and BUB1B promoter regions used for luciferase reporter assay. pSS-MAD2 construct contains an 1128-bp region (−1098 to +30 nt) of the MAD2 gene and the pSS-BUB1B construct contains a 1546-bp region (−1454 to +92 nt) of the BUB1B gene, cloned into pGL3 basic vector. MCF-7 cells were transiently transfected with pSS-MAD2 (250 ng) and pSS-BUB1B (500 ng), followed by E2 treatment, and proceeded for luciferase reporter assay. **p < 0.005; (two-tailed unpaired Student’s t-test) (C) Ectopic ERα up-regulates MAD2 and BUB1B promoter activity in MDA-MB-231 cells. MDA-MB-231 cells were co-transfected with pSS-MAD2 (250 ng) and pSS-BUB1B (500 ng) along with 1.5 µg of ectopic ERα (pCMV3-N-FLAG-ESR1), followed by E2 treatment and proceeded for luciferase reporter assay and plotted as discussed in (B). *p < 0.05; **p < 0.005; ***p < 0.0005 (two-tailed unpaired Student’s t-test). (D) Deletion of ERE results in a significant decrease in luciferase activity. Map of the two deletion constructs, ΔERE pSS-MAD2, and ΔERE pSS-BUB1B were shown in the top panel. MCF-7 cells were transfected with either ΔERE pSS-MAD2 (250 ng) or ΔERE pSS-BUB1B (500 ng), followed by E2 treatment. Data were plotted as discussed in (B). **p < 0.005; ns, not significant (two-tailed unpaired Student’s t-test). (E) ERα physically recruits to MAD2 and BUB1B promoters. The total cell extract of MCF-7 was prepared and Chromatin immunoprecipitation was done using antibodies specific for ERα and normal IgG as control. Precipitated chromatin was estimated by qRT-PCR. The results are expressed as a percent of input. *p < 0.05; **p < 0.005; (two-tailed unpaired Student’s t-test). (F) Representative scatter plot depicting the Pearson’s correlation analysis between the gene expression values (TPM) of MAD2 and BUB1B , extracted from TCGA BRCA cohort using UCSC Xena database. The Pearson correlation coefficient is reported as the ‘r’ value. ** *p < 0.0005; (two-tailed unpaired Student’s t-test). All the experiments (B-E) were performed in triplicates and error bars were represented as means ± s.d.

    Article Snippet: The fragments were then subcloned into luciferase reporter vector pGL3 basic (Promega) using restriction enzyme BglII (New England Biolabs).

    Techniques: Activity Assay, Luciferase, Reporter Assay, Construct, Clone Assay, Plasmid Preparation, Transfection, Two Tailed Test, Chromatin Immunoprecipitation, Control, Quantitative RT-PCR, Expressing