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pet28a tfam expression vector  (Addgene inc)


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    Addgene inc pet28a tfam expression vector
    Pet28a Tfam Expression Vector, supplied by Addgene inc, used in various techniques. Bioz Stars score: 94/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pet+expression+vector/pET-28a(%2B)-FGF2-K128N+(Plasmid+%23120284)/pm41730877-423-5-8
    Average 94 stars, based on 11 article reviews
    pet28a tfam expression vector - by Bioz Stars, 2026-09
    94/100 stars

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    Related Articles

    Plasmid Preparation:

    Article Title: Single-engineered-residue solvation perturbations regulate global protein architecture and function.
    Article Snippet: .. Each TAG-mutant pET28a-phoA plasmid was co-transformed with pEVOL-pAzF (Addgene, plasmid no. 31186) into Escherichia coli BL21(DE3) (Novagen). .. Cells were cultured in LB medium with 50 μg/mL kanamycin (SigmaAldrich) and 34 μg/mL chloramphenicol (Sigma-Aldrich) at 37 °C.

    Article Title: Genetically encoded actuators for acoustic manipulation
    Article Snippet: .. GV-expressing cells were produced by transforming a pET28a plasmid containing the bARG1 gene cluster (Addgene #106473) into BL21 (A1) E. coli (Thermo Fisher Scientific). ..

    Article Title: PRMT5 in mitochondria regulates mtDNA stability through TFAM arginine methylation
    Article Snippet: The reactions were stopped by adding 2 × SDS loading buffer (Invitrogen) and boiling the samples for 5 m. The resulting methylation products were separated by SDS–PAGE, transferred onto a PVDF membrane, and analyzed by Western blotting using an anti-SDMA antibody. .. TFAM purification was done using pET28a-TFAM expression vector (Addgene), which encompasses residues 43–246, representing the full-length TFAM protein as described previously .PRMT5 purification was done using the pET28a-PRMT5 expression vector.BL21 (DE3) E. coli was transformed with either pET28a-TFAM or pET28a-PRMT5 plasmids, and a single colony was inoculated in 20 ml of LB media containing 50 μg/ml Kanamycin and incubated for 12 h at 37 °C. ..

    Article Title: Defining AV2-1 as a novel pharmacological probe to target human and rodent TRPV2.
    Article Snippet: .. As previously published (Fluck et al., 2021; Haug et al., 2024; Pumroy et al., 2019; Pumroy et al., 2022; Pumroy et al., 2024; Rocereta et al., 2025), MSP2N2 was cloned into a pET28a vector (Addgene) and expressed in BL21 Escherichia coli. .. Cells were resuspended in lysis buffer (20-mM Tris-HCl, pH 8.0, 100-mM NaCl, 1-mM PMSF and complete EDTA-free protease inhibitor cocktail [Roche]) and mixed with 1-mM MgCl2, 1-mM CaCl2 and DNAse I (MilliporeSigma) for 15 min at 4 C to fully solubilise the cells.

    Article Title: The SETD2 L1609P mutation found in leukemia disrupts methyltransferase activity and reduces histone H3K36 trimethylation
    Article Snippet: .. A pet28a-MHL plasmid containing the cDNA coding for the human SETD2 catalytic domain (Addgene #25348, residues 1433–1711) was used in order to produce 6xHis-tagged WT SETD2 in BL21 HI-control (DE3) E . coli . .. SETD2 L1609P mutation was introduced using the QuickChange XL site-directed mutagenesis kit (Agilent) with the following primers: His_SETD2_L1609P_F: CACTATTACTTTATGGCTCCGAAAAATGATGAAATTATCGACGCG.

    Article Title: PRMT5 in mitochondria regulates mtDNA stability through TFAM arginine methylation.
    Article Snippet: The reactions were stopped by adding 2 × SDS loading buffer (Invitrogen) and boiling the samples for 5 m. The resulting methylation products were separated by SDS–PAGE, transferred onto a PVDF membrane, and analyzed by Western blotting using an anti-SDMA antibody. .. TFAM purification was done using pET28a-TFAM expression vector (Addgene) which encompasses residues 43–246, representing the full-length TFAM protein as described previously21.PRMT5 purification was done using pET28a-PRMT5 expression vector.BL21 (DE3) E. coli was transformed with eitherpET28a-TFAM or pET28a-PRMT5 plasmids and a single colony was inoculated in 20 ml of LB media containing 50 μg/ml Kanamycin and incubated for 12 h at 37°C. ..

    Article Title: BamA self-insertion drives inner-to-outer membrane remodelling in diderm Firmicutes.
    Article Snippet: .. Each protein was cloned into the multiple cloning site of the pET28a+ backbone (Addgene plasmid 2565) (Supplementary Table 9). ..

    Produced:

    Article Title: Genetically encoded actuators for acoustic manipulation
    Article Snippet: .. GV-expressing cells were produced by transforming a pET28a plasmid containing the bARG1 gene cluster (Addgene #106473) into BL21 (A1) E. coli (Thermo Fisher Scientific). ..

    Purification:

    Article Title: PRMT5 in mitochondria regulates mtDNA stability through TFAM arginine methylation
    Article Snippet: The reactions were stopped by adding 2 × SDS loading buffer (Invitrogen) and boiling the samples for 5 m. The resulting methylation products were separated by SDS–PAGE, transferred onto a PVDF membrane, and analyzed by Western blotting using an anti-SDMA antibody. .. TFAM purification was done using pET28a-TFAM expression vector (Addgene), which encompasses residues 43–246, representing the full-length TFAM protein as described previously .PRMT5 purification was done using the pET28a-PRMT5 expression vector.BL21 (DE3) E. coli was transformed with either pET28a-TFAM or pET28a-PRMT5 plasmids, and a single colony was inoculated in 20 ml of LB media containing 50 μg/ml Kanamycin and incubated for 12 h at 37 °C. ..

    Article Title: PRMT5 in mitochondria regulates mtDNA stability through TFAM arginine methylation.
    Article Snippet: The reactions were stopped by adding 2 × SDS loading buffer (Invitrogen) and boiling the samples for 5 m. The resulting methylation products were separated by SDS–PAGE, transferred onto a PVDF membrane, and analyzed by Western blotting using an anti-SDMA antibody. .. TFAM purification was done using pET28a-TFAM expression vector (Addgene) which encompasses residues 43–246, representing the full-length TFAM protein as described previously21.PRMT5 purification was done using pET28a-PRMT5 expression vector.BL21 (DE3) E. coli was transformed with eitherpET28a-TFAM or pET28a-PRMT5 plasmids and a single colony was inoculated in 20 ml of LB media containing 50 μg/ml Kanamycin and incubated for 12 h at 37°C. ..

    Expressing:

    Article Title: PRMT5 in mitochondria regulates mtDNA stability through TFAM arginine methylation
    Article Snippet: The reactions were stopped by adding 2 × SDS loading buffer (Invitrogen) and boiling the samples for 5 m. The resulting methylation products were separated by SDS–PAGE, transferred onto a PVDF membrane, and analyzed by Western blotting using an anti-SDMA antibody. .. TFAM purification was done using pET28a-TFAM expression vector (Addgene), which encompasses residues 43–246, representing the full-length TFAM protein as described previously .PRMT5 purification was done using the pET28a-PRMT5 expression vector.BL21 (DE3) E. coli was transformed with either pET28a-TFAM or pET28a-PRMT5 plasmids, and a single colony was inoculated in 20 ml of LB media containing 50 μg/ml Kanamycin and incubated for 12 h at 37 °C. ..

    Article Title: PRMT5 in mitochondria regulates mtDNA stability through TFAM arginine methylation.
    Article Snippet: The reactions were stopped by adding 2 × SDS loading buffer (Invitrogen) and boiling the samples for 5 m. The resulting methylation products were separated by SDS–PAGE, transferred onto a PVDF membrane, and analyzed by Western blotting using an anti-SDMA antibody. .. TFAM purification was done using pET28a-TFAM expression vector (Addgene) which encompasses residues 43–246, representing the full-length TFAM protein as described previously21.PRMT5 purification was done using pET28a-PRMT5 expression vector.BL21 (DE3) E. coli was transformed with eitherpET28a-TFAM or pET28a-PRMT5 plasmids and a single colony was inoculated in 20 ml of LB media containing 50 μg/ml Kanamycin and incubated for 12 h at 37°C. ..

    Transformation Assay:

    Article Title: PRMT5 in mitochondria regulates mtDNA stability through TFAM arginine methylation
    Article Snippet: The reactions were stopped by adding 2 × SDS loading buffer (Invitrogen) and boiling the samples for 5 m. The resulting methylation products were separated by SDS–PAGE, transferred onto a PVDF membrane, and analyzed by Western blotting using an anti-SDMA antibody. .. TFAM purification was done using pET28a-TFAM expression vector (Addgene), which encompasses residues 43–246, representing the full-length TFAM protein as described previously .PRMT5 purification was done using the pET28a-PRMT5 expression vector.BL21 (DE3) E. coli was transformed with either pET28a-TFAM or pET28a-PRMT5 plasmids, and a single colony was inoculated in 20 ml of LB media containing 50 μg/ml Kanamycin and incubated for 12 h at 37 °C. ..

    Article Title: PRMT5 in mitochondria regulates mtDNA stability through TFAM arginine methylation.
    Article Snippet: The reactions were stopped by adding 2 × SDS loading buffer (Invitrogen) and boiling the samples for 5 m. The resulting methylation products were separated by SDS–PAGE, transferred onto a PVDF membrane, and analyzed by Western blotting using an anti-SDMA antibody. .. TFAM purification was done using pET28a-TFAM expression vector (Addgene) which encompasses residues 43–246, representing the full-length TFAM protein as described previously21.PRMT5 purification was done using pET28a-PRMT5 expression vector.BL21 (DE3) E. coli was transformed with eitherpET28a-TFAM or pET28a-PRMT5 plasmids and a single colony was inoculated in 20 ml of LB media containing 50 μg/ml Kanamycin and incubated for 12 h at 37°C. ..

    Incubation:

    Article Title: PRMT5 in mitochondria regulates mtDNA stability through TFAM arginine methylation
    Article Snippet: The reactions were stopped by adding 2 × SDS loading buffer (Invitrogen) and boiling the samples for 5 m. The resulting methylation products were separated by SDS–PAGE, transferred onto a PVDF membrane, and analyzed by Western blotting using an anti-SDMA antibody. .. TFAM purification was done using pET28a-TFAM expression vector (Addgene), which encompasses residues 43–246, representing the full-length TFAM protein as described previously .PRMT5 purification was done using the pET28a-PRMT5 expression vector.BL21 (DE3) E. coli was transformed with either pET28a-TFAM or pET28a-PRMT5 plasmids, and a single colony was inoculated in 20 ml of LB media containing 50 μg/ml Kanamycin and incubated for 12 h at 37 °C. ..

    Article Title: PRMT5 in mitochondria regulates mtDNA stability through TFAM arginine methylation.
    Article Snippet: The reactions were stopped by adding 2 × SDS loading buffer (Invitrogen) and boiling the samples for 5 m. The resulting methylation products were separated by SDS–PAGE, transferred onto a PVDF membrane, and analyzed by Western blotting using an anti-SDMA antibody. .. TFAM purification was done using pET28a-TFAM expression vector (Addgene) which encompasses residues 43–246, representing the full-length TFAM protein as described previously21.PRMT5 purification was done using pET28a-PRMT5 expression vector.BL21 (DE3) E. coli was transformed with eitherpET28a-TFAM or pET28a-PRMT5 plasmids and a single colony was inoculated in 20 ml of LB media containing 50 μg/ml Kanamycin and incubated for 12 h at 37°C. ..

    Clone Assay:

    Article Title: Defining AV2-1 as a novel pharmacological probe to target human and rodent TRPV2.
    Article Snippet: .. As previously published (Fluck et al., 2021; Haug et al., 2024; Pumroy et al., 2019; Pumroy et al., 2022; Pumroy et al., 2024; Rocereta et al., 2025), MSP2N2 was cloned into a pET28a vector (Addgene) and expressed in BL21 Escherichia coli. .. Cells were resuspended in lysis buffer (20-mM Tris-HCl, pH 8.0, 100-mM NaCl, 1-mM PMSF and complete EDTA-free protease inhibitor cocktail [Roche]) and mixed with 1-mM MgCl2, 1-mM CaCl2 and DNAse I (MilliporeSigma) for 15 min at 4 C to fully solubilise the cells.

    Article Title: BamA self-insertion drives inner-to-outer membrane remodelling in diderm Firmicutes.
    Article Snippet: .. Each protein was cloned into the multiple cloning site of the pET28a+ backbone (Addgene plasmid 2565) (Supplementary Table 9). ..

    Cloning:

    Article Title: BamA self-insertion drives inner-to-outer membrane remodelling in diderm Firmicutes.
    Article Snippet: .. Each protein was cloned into the multiple cloning site of the pET28a+ backbone (Addgene plasmid 2565) (Supplementary Table 9). ..

    Construct:




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    Image Search Results


    Hypothesis and experiment system (A) We hypothesize that vertical and horizontal gene transfer (VGT and HGT) are influenced by the characteristics of the potential recipient cell types and determine the proliferation and diversity of transconjugant cells. Because the potential recipient community comprises multiple cell types with varying growth traits and conjugation probabilities, we expect the resulting composition of transconjugant cells to be shaped by these cell type-specific traits. (B) Our experimental system consists of E . coli MG1655 lacI q -pLpp-mCherry as the plasmid donor strain and pB10 as the focal plasmid. pB10 donor cells express RFP from the chromosome and transconjugants express GFP from pB10.

    Journal: iScience

    Article Title: Horizontal and vertical gene transfer shape the plasmid host range in surface-associated microbial systems

    doi: 10.1016/j.isci.2026.115299

    Figure Lengend Snippet: Hypothesis and experiment system (A) We hypothesize that vertical and horizontal gene transfer (VGT and HGT) are influenced by the characteristics of the potential recipient cell types and determine the proliferation and diversity of transconjugant cells. Because the potential recipient community comprises multiple cell types with varying growth traits and conjugation probabilities, we expect the resulting composition of transconjugant cells to be shaped by these cell type-specific traits. (B) Our experimental system consists of E . coli MG1655 lacI q -pLpp-mCherry as the plasmid donor strain and pB10 as the focal plasmid. pB10 donor cells express RFP from the chromosome and transconjugants express GFP from pB10.

    Article Snippet: MBP- mCherry expression plasmid (Amp R ) , Addgene , Plasmid# 29747.

    Techniques: Conjugation Assay, Plasmid Preparation

    Transconjugant proportions and diversities after surface-associated conjugation assays for different environmental conditions (A) Proportion of transconjugant cells relative to total cells after surface-associated conjugation assays using the WWTP community as the potential recipient cell population. We conducted conjugation assays on 1×SWW, 10×SWW, or LB agar plates using E . coli MG1655 lacI q -pLpp-mCherry as the pB10 donor strain. (B) Relative abundances of bacterial class in the total potential recipient cell population (T) and the transconjugant cell population (TC) as identified by 16S rRNA gene sequencing. We separated and identified TC cells using FC-FACS-sorting of GFP-positive cells. (C) Normalized Shannon index of the transconjugant populations after surface-associated conjugation assays on 1×SWW, 10×SWW, or LB agar plates. We normalized the Shannon index of the TC populations to their corresponding T populations. (D) Principal coordinate analysis (PCoA) based on weighted UniFrac distances of T and TC populations after surface-associated conjugation assays on 1×SWW, 10×SWW, or LB agar plates. (E) Phylogenetic tree of transconjugant ASVs detected after surface-associated conjugation assays on 1×SWW, 10×SWW, or LB agar plates. The outer colored box denotes the bacterial phylum of each ASV, corresponding to the phylum-level groupings shown in panel (B). The inner heatmap box aligned with each tip shows the log 10 fold-changes in ASV abundance (TC relative to T) across the three conditions. For (A and C), each point is an independent biological replicate ( n = 3), horizontal bars are the means, error bars are ±1 standard deviation, and asterisks indicate statistically significant differences between the means based on two-way ANOVA with Holm correction (∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, ns = not significant). For (D), each point is an independent biological replicate ( n = 3).

    Journal: iScience

    Article Title: Horizontal and vertical gene transfer shape the plasmid host range in surface-associated microbial systems

    doi: 10.1016/j.isci.2026.115299

    Figure Lengend Snippet: Transconjugant proportions and diversities after surface-associated conjugation assays for different environmental conditions (A) Proportion of transconjugant cells relative to total cells after surface-associated conjugation assays using the WWTP community as the potential recipient cell population. We conducted conjugation assays on 1×SWW, 10×SWW, or LB agar plates using E . coli MG1655 lacI q -pLpp-mCherry as the pB10 donor strain. (B) Relative abundances of bacterial class in the total potential recipient cell population (T) and the transconjugant cell population (TC) as identified by 16S rRNA gene sequencing. We separated and identified TC cells using FC-FACS-sorting of GFP-positive cells. (C) Normalized Shannon index of the transconjugant populations after surface-associated conjugation assays on 1×SWW, 10×SWW, or LB agar plates. We normalized the Shannon index of the TC populations to their corresponding T populations. (D) Principal coordinate analysis (PCoA) based on weighted UniFrac distances of T and TC populations after surface-associated conjugation assays on 1×SWW, 10×SWW, or LB agar plates. (E) Phylogenetic tree of transconjugant ASVs detected after surface-associated conjugation assays on 1×SWW, 10×SWW, or LB agar plates. The outer colored box denotes the bacterial phylum of each ASV, corresponding to the phylum-level groupings shown in panel (B). The inner heatmap box aligned with each tip shows the log 10 fold-changes in ASV abundance (TC relative to T) across the three conditions. For (A and C), each point is an independent biological replicate ( n = 3), horizontal bars are the means, error bars are ±1 standard deviation, and asterisks indicate statistically significant differences between the means based on two-way ANOVA with Holm correction (∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, ns = not significant). For (D), each point is an independent biological replicate ( n = 3).

    Article Snippet: MBP- mCherry expression plasmid (Amp R ) , Addgene , Plasmid# 29747.

    Techniques: Conjugation Assay, Sequencing, Standard Deviation

    Transconjugant growth during surface-associated conjugation assays for different environmental conditions (A) Representative fluorescence microscopy images of transconjugant cells during surface-associated conjugation assays on LB agar plates. E . coli MG1655 lacI q -pLpp-mCherry is the pB10 donor strain and show red fluorescence. Transconjugant cells are green. The time indicated in the images refers to the point at which transconjugant cells first became detectable. (B) Normalized microcolony area ( A / a 0 ) plotted as a function of time during the surface-associated conjugation assays on LB agar plates. A is the total microcolony area and a 0 is the initial transconjugant area. Connected data points are for individual colonies ( n = 12). (C) Microcolony area at the endpoint of the mating assay (t = 24 h) for different environmental conditions. The half-violin and scatterplots present the sample distribution and individual microcolony measurements for surface-associated conjugation assays on different medium (n 1xSWW = 880, n 10xSWW = 664, n LB = 1,070, for microcolony number). We performed each experiment at least three independent experiments. Horizontal bars are the mean microcolony areas, error bars are the 99% confidence intervals, and asterisks indicate statistically significant differences between the means based on two-way ANOVA with Holm correction (∗∗ p < 0.01, ∗∗∗∗ p < 0.0001, ns = not significant).

    Journal: iScience

    Article Title: Horizontal and vertical gene transfer shape the plasmid host range in surface-associated microbial systems

    doi: 10.1016/j.isci.2026.115299

    Figure Lengend Snippet: Transconjugant growth during surface-associated conjugation assays for different environmental conditions (A) Representative fluorescence microscopy images of transconjugant cells during surface-associated conjugation assays on LB agar plates. E . coli MG1655 lacI q -pLpp-mCherry is the pB10 donor strain and show red fluorescence. Transconjugant cells are green. The time indicated in the images refers to the point at which transconjugant cells first became detectable. (B) Normalized microcolony area ( A / a 0 ) plotted as a function of time during the surface-associated conjugation assays on LB agar plates. A is the total microcolony area and a 0 is the initial transconjugant area. Connected data points are for individual colonies ( n = 12). (C) Microcolony area at the endpoint of the mating assay (t = 24 h) for different environmental conditions. The half-violin and scatterplots present the sample distribution and individual microcolony measurements for surface-associated conjugation assays on different medium (n 1xSWW = 880, n 10xSWW = 664, n LB = 1,070, for microcolony number). We performed each experiment at least three independent experiments. Horizontal bars are the mean microcolony areas, error bars are the 99% confidence intervals, and asterisks indicate statistically significant differences between the means based on two-way ANOVA with Holm correction (∗∗ p < 0.01, ∗∗∗∗ p < 0.0001, ns = not significant).

    Article Snippet: MBP- mCherry expression plasmid (Amp R ) , Addgene , Plasmid# 29747.

    Techniques: Conjugation Assay, Fluorescence, Microscopy