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empty pgl3 c vector (New England Biolabs)

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New England Biolabs empty pgl3 c vector
Empty Pgl3 C Vector, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 9085 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/pgl3+empty+vector/XhoI/pm38740892-243-6-32
Average 99 stars, based on 9085 article reviews

empty pgl3 c vector - by Bioz Stars, 2026-10

99/100 stars

Images

Plasmid Preparation:

Article Title: CelFDrive: Multimodal deep learning-assisted microscopy for automated detection of rare events.
Article Snippet: For live-cell DNA visualisation, 0.5 μM SiR-DNA (Spirochrome) was added to the cell-culture medium at least 1 h before imaging. .. Plasmid construction To generate the 3×mScarlet3–α-tubulin plasmid, cDNA encoding three repeats of mScarlet3 (Gadella et al., 2023) was synthesised by GeneArt (Life Technologies) and subcloned into the N-terminal region of a plasmid expressing α-tubulin (Addgene plasmid 189768) using NheI and XhoI restriction enzymes (NEB). .. The plasmid encoding HALO-NUP107 for expression in mammalian cells was synthesised by GeneArt (Life Technologies).

Article Title: Next‐Generation Sequencing to Identify Genetic Causes of Chinese Patients Affected With Non‐retinal Developmental Eye Disorders
Article Snippet: .. Plasmid pSPL3 served as the plasmid backbone and was digested using two restriction enzymes, XhoI and BamHI (New England Biolabs, Ipswich, Massachusetts, USA). ..

Article Title: Injectable mRNA-engineered short fibers deliver temporally coupled mechano-genetic signals to reprogram Schwann cell cytoskeleton for nerve repair.
Article Snippet: To eliminate endotoxin contamination, plasmid templates were isolated using FastPure Enhanced EndoFree Plasmid Maxi Kit (DC212, Vazyme, Nanjing, China), guaranteeing endotoxin levels<0.1 EU/μg. .. The plasmid was linearized using the restriction enzyme XhoI (R0146M, New England Biolabs, Ipswich, MA) and transcribed into mRNA using T7 High Yield RNA Transcription Kit (E131, Novoprotein, Suzhou, China). ..

Article Title: Tryptophan oxidation-induced dynamics highlight why stable γS-crystallin variants aggregate.
Article Snippet: The gene fragment for His-tagged γS-W163S (Sup- 121 plementary Figure S1) was purchased from Twist Bioscience (South San Francisco, California). .. 122 Restriction enzymes NcoI and XhoI and Gibson assembly master mix were purchased from New 123 England Biolabs (Ipswich, Massachusetts) and used to assemble the pET28a(+) vector contain- 124 ing the gene for γS-W163S. .. Genes of interest within plasmid samples were verified by Sanger 125 sequencing using the T7 universal primer provided by GENEWIZ from Azenta (South Plainfield, 126 New Jersey).

Article Title: Sustained Relief of Chronic Pain via a Na v 1.7-Targeting ASO-siRNA Conjugate in Rats.
Article Snippet: 576 577 psiCHECK assay 578 Off-target activity was assessed using the Dual-Glo Luciferase Assay System (Promega, 579 Cat. E2920). .. The psiCHECK-2 vector contains a constitutively expressed firefly luciferase 580 cassette and a multiple cloning site located in the 3’-UTR of the Renilla luciferase gene.80 581 Oligonucleotides containing three tandem repeats of seed-matched (SM) sequences (Table 582 S5) were synthesized with XhoI (New England Biolabs, Cat. R0146S) and NotI (New 583 England Biolabs, Cat. R3189S) restriction sites at the ends. ..

Article Title: MaltEms-MaltOBP9 axis mediates sexually dimorphic olfactory behavior in Monochamus alternatus via recognition of pheromone 2-methoxy-4-vinylphenol.
Article Snippet: The olfactory system is essential for insect survival and behavior, with odorant-binding proteins (OBPs) playing a key role by binding and transporting chemical signals to olfactory receptors.. Although MaltOBP9 from Monochamus alternatus, a primary vector of pine wilt disease, has been linked to olfactory functions in M. alternatus, its regulatory mechanisms and specific roles are not well understood.. In this study, we show that MaltOBP9 is highly expressed in the antennae of both male and female adults after sexual maturation.

Expressing:

Polymerase Chain Reaction:

Article Title: Cryo-EM structure of a methanogen nitrogenase-PII protein supercomplex.
Article Snippet: .. The PCR product was digested with NcoI and XhoI according to the manufacturer’s instructions (NEB) and ligated into NcoI/XhoI-digested pET28a using T4 DNA ligase with incubation for 16 h at 16 °C. ..

Incubation:

other:

Article Title: The ROCK1 PHC1 domain mediates recruitment to Rho activity zones and contractility signaling.
Article Snippet: The resulting plasmid was used as a backbone for the Gibson assembly after restriction digestion using XhoI (NEB).

Luciferase:

Cloning:

Synthesized:

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

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