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pgl3 basic luciferase reporter vector  (Addgene inc)


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    Structured Review

    Addgene inc pgl3 basic luciferase reporter vector
    Pgl3 Basic Luciferase Reporter Vector, supplied by Addgene inc, used in various techniques. Bioz Stars score: 95/100, based on 195 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pgl3+basic+luciferase+reporter+vector/pGL3+(-BRE)+Luciferase+(Plasmid+%2345127)/pm39265509-80-14-18
    Average 95 stars, based on 195 article reviews
    pgl3 basic luciferase reporter vector - by Bioz Stars, 2026-09
    95/100 stars

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    Related Articles

    Clone Assay:

    Article Title: MG53 suppresses tumor growth via transcriptional inhibition of KIF11 in pancreatic cancer.
    Article Snippet: .. A 908-base pair fragment of the KIF11 promoter was cloned and inserted into the pGL3-basic luciferase reporter vector (RRID:Addgene_11,994), resulting in the creation of the KIF11 promoter luciferase reporter plasmid. ..

    Article Title: lncMGPF is a novel positive regulator of muscle growth and regeneration.
    Article Snippet: .. lncMGPF promoter regions were cloned into a PGL3‐basic luciferase reporter vector (Addgene, USA), and the constructed luciferase reporter vectors were transfected into C2C12 myoblasts in a 24‐well plate. .. The 3′ untranslated region (UTR) of MEF2C, sequences containing the miR‐135a‐5p binding site of lncMGPF or AK394747, and the sequences of lncMGPF or AK394747 with mutated miR‐135a‐5p binding sites, were cloned into the pmirGLO Dual‐Luciferase miRNA Target Expression Vector (Promega, USA) to construct the corresponding reporter vectors.

    Article Title: lncMGPF is a novel positive regulator of muscle growth and regeneration
    Article Snippet: .. lncMGPF promoter regions were cloned into a PGL3‐basic luciferase reporter vector (Addgene, USA), and the constructed luciferase reporter vectors were transfected into C2C12 myoblasts in a 24‐well plate. .. The 3′ untranslated region (UTR) of MEF2C , sequences containing the miR‐135a‐5p binding site of lncMGPF or AK394747 , and the sequences of lncMGPF or AK394747 with mutated miR‐135a‐5p binding sites, were cloned into the pmirGLO Dual‐Luciferase miRNA Target Expression Vector (Promega, USA) to construct the corresponding reporter vectors.

    Article Title: LncEDCH1 improves mitochondrial function to reduce muscle atrophy by interacting with SERCA2
    Article Snippet: .. Different fragments of the LncEDCH1 promoter were cloned into a pGL3-basic luciferase reporter vector (Addgene, USA), and the constructed luciferase reporter vectors were transfected into CPMs in a 96-well plate. .. The luciferase assay was performed using the Dual-Luciferase Reporter Assay System (Promega, USA) following the manufacturer's instructions.

    Luciferase:

    Article Title: MG53 suppresses tumor growth via transcriptional inhibition of KIF11 in pancreatic cancer.
    Article Snippet: .. A 908-base pair fragment of the KIF11 promoter was cloned and inserted into the pGL3-basic luciferase reporter vector (RRID:Addgene_11,994), resulting in the creation of the KIF11 promoter luciferase reporter plasmid. ..

    Article Title: lncMGPF is a novel positive regulator of muscle growth and regeneration.
    Article Snippet: .. lncMGPF promoter regions were cloned into a PGL3‐basic luciferase reporter vector (Addgene, USA), and the constructed luciferase reporter vectors were transfected into C2C12 myoblasts in a 24‐well plate. .. The 3′ untranslated region (UTR) of MEF2C, sequences containing the miR‐135a‐5p binding site of lncMGPF or AK394747, and the sequences of lncMGPF or AK394747 with mutated miR‐135a‐5p binding sites, were cloned into the pmirGLO Dual‐Luciferase miRNA Target Expression Vector (Promega, USA) to construct the corresponding reporter vectors.

    Article Title: lncMGPF is a novel positive regulator of muscle growth and regeneration
    Article Snippet: .. lncMGPF promoter regions were cloned into a PGL3‐basic luciferase reporter vector (Addgene, USA), and the constructed luciferase reporter vectors were transfected into C2C12 myoblasts in a 24‐well plate. .. The 3′ untranslated region (UTR) of MEF2C , sequences containing the miR‐135a‐5p binding site of lncMGPF or AK394747 , and the sequences of lncMGPF or AK394747 with mutated miR‐135a‐5p binding sites, were cloned into the pmirGLO Dual‐Luciferase miRNA Target Expression Vector (Promega, USA) to construct the corresponding reporter vectors.

    Article Title: LncEDCH1 improves mitochondrial function to reduce muscle atrophy by interacting with SERCA2
    Article Snippet: .. Different fragments of the LncEDCH1 promoter were cloned into a pGL3-basic luciferase reporter vector (Addgene, USA), and the constructed luciferase reporter vectors were transfected into CPMs in a 96-well plate. .. The luciferase assay was performed using the Dual-Luciferase Reporter Assay System (Promega, USA) following the manufacturer's instructions.

    Plasmid Preparation:

    Article Title: MG53 suppresses tumor growth via transcriptional inhibition of KIF11 in pancreatic cancer.
    Article Snippet: .. A 908-base pair fragment of the KIF11 promoter was cloned and inserted into the pGL3-basic luciferase reporter vector (RRID:Addgene_11,994), resulting in the creation of the KIF11 promoter luciferase reporter plasmid. ..

    Construct:

    Article Title: lncMGPF is a novel positive regulator of muscle growth and regeneration.
    Article Snippet: .. lncMGPF promoter regions were cloned into a PGL3‐basic luciferase reporter vector (Addgene, USA), and the constructed luciferase reporter vectors were transfected into C2C12 myoblasts in a 24‐well plate. .. The 3′ untranslated region (UTR) of MEF2C, sequences containing the miR‐135a‐5p binding site of lncMGPF or AK394747, and the sequences of lncMGPF or AK394747 with mutated miR‐135a‐5p binding sites, were cloned into the pmirGLO Dual‐Luciferase miRNA Target Expression Vector (Promega, USA) to construct the corresponding reporter vectors.

    Article Title: lncMGPF is a novel positive regulator of muscle growth and regeneration
    Article Snippet: .. lncMGPF promoter regions were cloned into a PGL3‐basic luciferase reporter vector (Addgene, USA), and the constructed luciferase reporter vectors were transfected into C2C12 myoblasts in a 24‐well plate. .. The 3′ untranslated region (UTR) of MEF2C , sequences containing the miR‐135a‐5p binding site of lncMGPF or AK394747 , and the sequences of lncMGPF or AK394747 with mutated miR‐135a‐5p binding sites, were cloned into the pmirGLO Dual‐Luciferase miRNA Target Expression Vector (Promega, USA) to construct the corresponding reporter vectors.

    Article Title: LncEDCH1 improves mitochondrial function to reduce muscle atrophy by interacting with SERCA2
    Article Snippet: .. Different fragments of the LncEDCH1 promoter were cloned into a pGL3-basic luciferase reporter vector (Addgene, USA), and the constructed luciferase reporter vectors were transfected into CPMs in a 96-well plate. .. The luciferase assay was performed using the Dual-Luciferase Reporter Assay System (Promega, USA) following the manufacturer's instructions.

    Transfection:

    Article Title: lncMGPF is a novel positive regulator of muscle growth and regeneration.
    Article Snippet: .. lncMGPF promoter regions were cloned into a PGL3‐basic luciferase reporter vector (Addgene, USA), and the constructed luciferase reporter vectors were transfected into C2C12 myoblasts in a 24‐well plate. .. The 3′ untranslated region (UTR) of MEF2C, sequences containing the miR‐135a‐5p binding site of lncMGPF or AK394747, and the sequences of lncMGPF or AK394747 with mutated miR‐135a‐5p binding sites, were cloned into the pmirGLO Dual‐Luciferase miRNA Target Expression Vector (Promega, USA) to construct the corresponding reporter vectors.

    Article Title: lncMGPF is a novel positive regulator of muscle growth and regeneration
    Article Snippet: .. lncMGPF promoter regions were cloned into a PGL3‐basic luciferase reporter vector (Addgene, USA), and the constructed luciferase reporter vectors were transfected into C2C12 myoblasts in a 24‐well plate. .. The 3′ untranslated region (UTR) of MEF2C , sequences containing the miR‐135a‐5p binding site of lncMGPF or AK394747 , and the sequences of lncMGPF or AK394747 with mutated miR‐135a‐5p binding sites, were cloned into the pmirGLO Dual‐Luciferase miRNA Target Expression Vector (Promega, USA) to construct the corresponding reporter vectors.

    Article Title: LncEDCH1 improves mitochondrial function to reduce muscle atrophy by interacting with SERCA2
    Article Snippet: .. Different fragments of the LncEDCH1 promoter were cloned into a pGL3-basic luciferase reporter vector (Addgene, USA), and the constructed luciferase reporter vectors were transfected into CPMs in a 96-well plate. .. The luciferase assay was performed using the Dual-Luciferase Reporter Assay System (Promega, USA) following the manufacturer's instructions.



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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