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pgl3 promega empty firefly luciferase vector rc203227 origene denr  (Addgene inc)


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    Addgene inc pgl3 promega empty firefly luciferase vector rc203227 origene denr
    Pgl3 Promega Empty Firefly Luciferase Vector Rc203227 Origene Denr, supplied by Addgene inc, used in various techniques. Bioz Stars score: 94/100, based on 71 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pgl3+empty+vector/JDS246+(Plasmid+%2343861)/pm37862172-189-34-30
    Average 94 stars, based on 71 article reviews
    pgl3 promega empty firefly luciferase vector rc203227 origene denr - by Bioz Stars, 2026-11
    94/100 stars

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    Related Articles

    Plasmid Preparation:

    Article Title: Characterization and inactivation of endogenous retroviruses in chinese hamster ovary cells
    Article Snippet: .. Plasmid Construction The mammalian codon-optimized Streptococcus pyogenes Cas9 (SpCas9) nuclease expression plasmid JDS246 (ADDGENE plasmid #43861) (21) was used to introduce site-specific DSBs. ..

    Article Title: Cas9 variants and uses thereof
    Article Snippet: PCR was performed with 0.4 μL of 2 U/μL Phusion Hot Start Flex DNA polymerase (NEB) in 50 μL with 1×HF Buffer, 0.2 mM dNTP mix (0.2 mM dATP, 0.2 mM dCTP, 0.2 mM dGTP, 0.2 mM dTTP) (NEB), 0.5 μM of each primer and a program of: 98° C., 1 min; 35 cycles of [98° C. 15 s; 65° C., 15 s; 72° C., 30 s] unless otherwise noted. .. Construction of FokI-dCas9, Cas9 Nickase and gRNA Expression Plasmids The human codon-optimized Streptococcus pyogenes Cas9 nuclease with NLS and 3×FLAG tag (Addgene plasmid 43861) 2 was used as the wild-type Cas9 expression plasmid. .. PCR (72° C., 3 min) products of wild-type Cas9 expression plasmid as template with Cas9_Exp primers listed in Table 1 below were assembled with Gibson Assembly Cloning Kit (New England Biolabs) to construct Cas9 and FokI-dCas9 variants.

    Article Title: CAS9 proteins including ligand-dependent inteins
    Article Snippet: .. A plasmid encoding the human codon-optimized Streptococcus pyogenes Cas9 nuclease with an NLS and 3×FLAG tag (Addgene plasmid 43861)5 was used as the wild-type Cas9 expression plasmid. .. Intein 37R3-2 was subcloned at the described positions into the wild-type Cas9 expression plasmid using USER (NEB M5505) cloning. sgRNA expression plasmids used in this study have been described previously11.

    Article Title: <i>Giardia's</i> domed ventral disc architecture is essential for attachment and contributes to epithelial barrier disruption
    Article Snippet: Giardia lamblia is a widespread anaerobic protistan parasite causing significant diarrheal disease worldwide.. Giardia trophozoites attach extracellularly to the host gastrointestinal epithelium using a unique microtubule organelle, the ventral disc.. The complex, dome-shaped disc is composed of microribbon-crossbridge (MR-CB) protein complexes scaffolded onto a spiral microtubule array.

    Article Title: Inhibitors of RNA-guided nuclease target binding and uses thereof
    Article Snippet: In the plasmid nucleofection method, cells were nucleofected either with 440 ng of SpCas9 (Addgene Plasmid #43861) or 400 ng of SpCas9 along with 40 ng of sgRNA (pFYF1320 eGFP Site #1, Addgene Plasmid #47511). .. In the plasmid nucleofection method, cells were nucleofected either with 440 ng of SpCas9 (Addgene Plasmid #43861) or 400 ng of SpCas9 along with 40 ng of sgRNA (pFYF1320 eGFP Site #1, Addgene Plasmid #47511). ..

    Expressing:

    Article Title: Characterization and inactivation of endogenous retroviruses in chinese hamster ovary cells
    Article Snippet: .. Plasmid Construction The mammalian codon-optimized Streptococcus pyogenes Cas9 (SpCas9) nuclease expression plasmid JDS246 (ADDGENE plasmid #43861) (21) was used to introduce site-specific DSBs. ..

    Article Title: Cas9 variants and uses thereof
    Article Snippet: PCR was performed with 0.4 μL of 2 U/μL Phusion Hot Start Flex DNA polymerase (NEB) in 50 μL with 1×HF Buffer, 0.2 mM dNTP mix (0.2 mM dATP, 0.2 mM dCTP, 0.2 mM dGTP, 0.2 mM dTTP) (NEB), 0.5 μM of each primer and a program of: 98° C., 1 min; 35 cycles of [98° C. 15 s; 65° C., 15 s; 72° C., 30 s] unless otherwise noted. .. Construction of FokI-dCas9, Cas9 Nickase and gRNA Expression Plasmids The human codon-optimized Streptococcus pyogenes Cas9 nuclease with NLS and 3×FLAG tag (Addgene plasmid 43861) 2 was used as the wild-type Cas9 expression plasmid. .. PCR (72° C., 3 min) products of wild-type Cas9 expression plasmid as template with Cas9_Exp primers listed in Table 1 below were assembled with Gibson Assembly Cloning Kit (New England Biolabs) to construct Cas9 and FokI-dCas9 variants.

    Article Title: CAS9 proteins including ligand-dependent inteins
    Article Snippet: .. A plasmid encoding the human codon-optimized Streptococcus pyogenes Cas9 nuclease with an NLS and 3×FLAG tag (Addgene plasmid 43861)5 was used as the wild-type Cas9 expression plasmid. .. Intein 37R3-2 was subcloned at the described positions into the wild-type Cas9 expression plasmid using USER (NEB M5505) cloning. sgRNA expression plasmids used in this study have been described previously11.

    Introduce:

    Article Title: Characterization and inactivation of endogenous retroviruses in chinese hamster ovary cells
    Article Snippet: .. Plasmid Construction The mammalian codon-optimized Streptococcus pyogenes Cas9 (SpCas9) nuclease expression plasmid JDS246 (ADDGENE plasmid #43861) (21) was used to introduce site-specific DSBs. ..

    other:

    Article Title: Unconstrained genome targeting with near-PAMless engineered CRISPR-Cas9 variants
    Article Snippet: Cytidine base editor (CBE) constructs were generated by subcloning the open reading frame of BE4max (Addgene plasmid 112099; a gift from David Liu) into the NotI and AgeI sites of pCAG-CFP (Addgene plasmid 11179; a gift from Connie Cepko).

    Article Title: Inhibitors of RNA-guided nuclease target binding and uses thereof
    Article Snippet: NA Software and Algorithms GraphPad Prism GraphPad Prism Software NA Inc. Spotfire Tibco NA MetaXpress Molecular Device NA MATLAB MathWorks NA



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    (a) Schematic diagram for constructing wild-type or mutant promoter at the ABCG1 rs57137919G>A polymorphism site. Two constructs were subcloned into <t>pGL3-basic</t> plasmid vectors with firefly luciferase reporter gene. pG, -367G (open bar); pA, -367A (grey bar). Luciferase activity assays for two ABCG1 promoter constructs in HEK293T cells (b), THP-1 cells (c), and HepG2 cells (d) are shown. Luciferase activities were measured with the dual-luciferase reporter assay system and normalized to Renilla luciferase activity 24 hours after transfection in the presence or absence of TO901317. The luciferase activity was a corrected relative value. Data are shown as mean ± SD of four independent experiments in triplicate. P <0.01 vs. pG in HEK293 cells, THP-1 cells, and HepG2 cells under both basal state and LXR agonist stimulation.
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    Image Search Results


    (A.i.) Giemsa-stained images of LD-S, LD-R, in murine peritoneal MФs with or without the presence of suboptimal dose of SbV (1.52µg/ml). In some experimental condition LD infection was performed in the presence of N-acetyl-L-cysteine (NAC), while in some experimental conditions, LDs are pre-treated with H 2 O 2 + 6-AN. All the infections were performed for 24hrs (A.ii.) Bar graph showing amastigotes/100MФs in all the experimental sets mentioned in A.i. (B.) Western blot of whole cell lysate showing significant high expression of Heme-oxygenase 1 (HO1) in LD-R infected MФs while no significant change in expression level was observed for Ferritin (FTH1) at 4hrs pi. β-actin is used as the positive control. (C.) Schematic representation of HO-1 promoter region with p50 and c-Rel binding sites. Site A (−4/−635) was demarcated as green, and Site B (−636/−1377) was demarcated as red. (D.i.) Western blot of nuclear fraction showing significant high expression of p50 and c-Rel in LD-R infected MФs 4hrs pi with Histone (H3) as loading control. (D.ii.) Confocal images representing the localization of p50 and c-Rel, in MФs, with DAPI representing the nucleus. The right-most panel shows the RGB-profile plot with grey regions demarcating the region where p50 and cRel are colocalized with DAPI. p50 and c-Rel were found to be localized in the nucleus of LD-R-infected MФs. One representative small nucleus of LD has been marked in ( * ) to show the infected MФs. (E.) Fold luciferase activity of RAW264.7 cell lysate transfected either with HO1+PGL3 promoter, or Site A ( −/− ), Site B( −/− ) deleted constructs followed by infection with LD-S and LD-R for 4hrs. PGL3 enhancer empty vector is used for normalization for each transfected set. Each experiment was performed in 3 biological replicates and graphical data are represented as Mean with SEM. P ≤ 0.05 is marked as *, P ≤ 0.01 is marked as **, P ≤ 0.001 is marked as ***, and P ≤ 0.0001 is marked as ****.

    Journal: bioRxiv

    Article Title: Addressing anemia severity in antimony-resistant Leishmania donovani infection at the nexus of oxidative outburst and iron transit

    doi: 10.1101/2024.03.04.583250

    Figure Lengend Snippet: (A.i.) Giemsa-stained images of LD-S, LD-R, in murine peritoneal MФs with or without the presence of suboptimal dose of SbV (1.52µg/ml). In some experimental condition LD infection was performed in the presence of N-acetyl-L-cysteine (NAC), while in some experimental conditions, LDs are pre-treated with H 2 O 2 + 6-AN. All the infections were performed for 24hrs (A.ii.) Bar graph showing amastigotes/100MФs in all the experimental sets mentioned in A.i. (B.) Western blot of whole cell lysate showing significant high expression of Heme-oxygenase 1 (HO1) in LD-R infected MФs while no significant change in expression level was observed for Ferritin (FTH1) at 4hrs pi. β-actin is used as the positive control. (C.) Schematic representation of HO-1 promoter region with p50 and c-Rel binding sites. Site A (−4/−635) was demarcated as green, and Site B (−636/−1377) was demarcated as red. (D.i.) Western blot of nuclear fraction showing significant high expression of p50 and c-Rel in LD-R infected MФs 4hrs pi with Histone (H3) as loading control. (D.ii.) Confocal images representing the localization of p50 and c-Rel, in MФs, with DAPI representing the nucleus. The right-most panel shows the RGB-profile plot with grey regions demarcating the region where p50 and cRel are colocalized with DAPI. p50 and c-Rel were found to be localized in the nucleus of LD-R-infected MФs. One representative small nucleus of LD has been marked in ( * ) to show the infected MФs. (E.) Fold luciferase activity of RAW264.7 cell lysate transfected either with HO1+PGL3 promoter, or Site A ( −/− ), Site B( −/− ) deleted constructs followed by infection with LD-S and LD-R for 4hrs. PGL3 enhancer empty vector is used for normalization for each transfected set. Each experiment was performed in 3 biological replicates and graphical data are represented as Mean with SEM. P ≤ 0.05 is marked as *, P ≤ 0.01 is marked as **, P ≤ 0.001 is marked as ***, and P ≤ 0.0001 is marked as ****.

    Article Snippet: Murine HO-1 promoters (−1385/+137), 1522bp using primers 5’-AAGGTACCTGAGGCTGGAGAGATGGCC-3’ and 3’-TAAAAGCTTCACCGGACTGGGCTAGTTCAG-5’ were PCR amplified and cloned in promoterless PGL3 enhancer empty vector (Promega, E1771) at the upstream of luciferase gene.

    Techniques: Staining, Infection, Western Blot, Expressing, Positive Control, Binding Assay, Luciferase, Activity Assay, Transfection, Construct, Plasmid Preparation

    (a) Schematic diagram for constructing wild-type or mutant promoter at the ABCG1 rs57137919G>A polymorphism site. Two constructs were subcloned into pGL3-basic plasmid vectors with firefly luciferase reporter gene. pG, -367G (open bar); pA, -367A (grey bar). Luciferase activity assays for two ABCG1 promoter constructs in HEK293T cells (b), THP-1 cells (c), and HepG2 cells (d) are shown. Luciferase activities were measured with the dual-luciferase reporter assay system and normalized to Renilla luciferase activity 24 hours after transfection in the presence or absence of TO901317. The luciferase activity was a corrected relative value. Data are shown as mean ± SD of four independent experiments in triplicate. P <0.01 vs. pG in HEK293 cells, THP-1 cells, and HepG2 cells under both basal state and LXR agonist stimulation.

    Journal: PLoS ONE

    Article Title: ABCG1 rs57137919G>A Polymorphism Is Functionally Associated with Varying Gene Expression and Apoptosis of Macrophages

    doi: 10.1371/journal.pone.0097044

    Figure Lengend Snippet: (a) Schematic diagram for constructing wild-type or mutant promoter at the ABCG1 rs57137919G>A polymorphism site. Two constructs were subcloned into pGL3-basic plasmid vectors with firefly luciferase reporter gene. pG, -367G (open bar); pA, -367A (grey bar). Luciferase activity assays for two ABCG1 promoter constructs in HEK293T cells (b), THP-1 cells (c), and HepG2 cells (d) are shown. Luciferase activities were measured with the dual-luciferase reporter assay system and normalized to Renilla luciferase activity 24 hours after transfection in the presence or absence of TO901317. The luciferase activity was a corrected relative value. Data are shown as mean ± SD of four independent experiments in triplicate. P <0.01 vs. pG in HEK293 cells, THP-1 cells, and HepG2 cells under both basal state and LXR agonist stimulation.

    Article Snippet: The pGL3-basic empty vector was used as a negative control and the pGL3-control vector (Promega) was used as a positive control for the luciferase assay.

    Techniques: Mutagenesis, Construct, Plasmid Preparation, Luciferase, Activity Assay, Reporter Assay, Transfection