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length wt gfp pfkm construct  (Sino Biological)


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    Structured Review

    Sino Biological length wt gfp pfkm construct
    Length Wt Gfp Pfkm Construct, supplied by Sino Biological, used in various techniques. Bioz Stars score: 94/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/wt+construct/Human+PFK1%2FPFKM+Gene+ORF+cDNA+clone+expression+plasmid%2C+N-GFPSpark+tag/pmc12945692-446-1-7
    Average 94 stars, based on 2 article reviews
    length wt gfp pfkm construct - by Bioz Stars, 2026-10
    94/100 stars

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    Related Articles

    other:

    Article Title: PFKM governs metabolic shifts throughout skeletal muscle differentiation
    Article Snippet: Fiji (v.1.54p) (ImageJ, NIH) was used for blot analyses.

    Construct:

    Article Title: Functional Analyses of SATB2 Variants Reveal Pathogenicity Mechanisms Linked With SATB2-Associated Syndrome.
    Article Snippet: .. SATB2 proteins with the seven variants, including variants from previous reports with detailed descriptions of the oral phenotype (Kikuiri et al. 2018; Kurosaka et al. 2023; Li, Ye, and Su 2022), were constructed using expression with a wildtype (WT) SATB2 plasmid (Human SATB2 Gene ORF cDNA clone expression plasmid, N- Flag tag; Sino Biological Inc., Kanagawa, Japan) as the DNA template, using the QuickChange Lightening Site- directed Mutagenesis Kit (Agilent Technologies, La Jolla, CA, USA) according to the manufacturer's protocol. .. Variants were confirmed using the Applied Biosystems Big Dye Terminator Reaction kit and Applied Biosystems 3130xl Genetic Analyzer.

    Article Title: PFKM governs metabolic shifts throughout skeletal muscle differentiation.
    Article Snippet: .. The full-length WT-GFP–PFKM construct was purchased from SinoBiological (HG14133-ANG). ..

    Article Title: Activation and Purification of ß-Glucocerebrosidase by Exploiting its Transporter LIMP-2 - Implications for Novel Treatment Strategies in Gaucher's and Parkinson's Disease.
    Article Snippet: All other expression constructs used for mammalian expression were based on the pCMV3 vector (Sino Biological Inc., Peking, China, #CV011). .. For expression of fulllength human LIMP-2 (Sino Biological Inc., Peking, China, #HG11063CH), and untagged human GCase wt (Sino Biological Inc., Peking, China, #HG12038-UT), commercially available expression constructs were used. .. For the expression of 10xHis-tagged GCase wt, a commercially available plasmid (Sino Biological Inc., Peking, China, #HG12038-CH) was modified to make the C-terminal His tag cleavable by tobacco etch virus (TEV) protease.

    Expressing:

    Article Title: Functional Analyses of SATB2 Variants Reveal Pathogenicity Mechanisms Linked With SATB2-Associated Syndrome.
    Article Snippet: .. SATB2 proteins with the seven variants, including variants from previous reports with detailed descriptions of the oral phenotype (Kikuiri et al. 2018; Kurosaka et al. 2023; Li, Ye, and Su 2022), were constructed using expression with a wildtype (WT) SATB2 plasmid (Human SATB2 Gene ORF cDNA clone expression plasmid, N- Flag tag; Sino Biological Inc., Kanagawa, Japan) as the DNA template, using the QuickChange Lightening Site- directed Mutagenesis Kit (Agilent Technologies, La Jolla, CA, USA) according to the manufacturer's protocol. .. Variants were confirmed using the Applied Biosystems Big Dye Terminator Reaction kit and Applied Biosystems 3130xl Genetic Analyzer.

    Article Title: Activation and Purification of ß-Glucocerebrosidase by Exploiting its Transporter LIMP-2 - Implications for Novel Treatment Strategies in Gaucher's and Parkinson's Disease.
    Article Snippet: All other expression constructs used for mammalian expression were based on the pCMV3 vector (Sino Biological Inc., Peking, China, #CV011). .. For expression of fulllength human LIMP-2 (Sino Biological Inc., Peking, China, #HG11063CH), and untagged human GCase wt (Sino Biological Inc., Peking, China, #HG12038-UT), commercially available expression constructs were used. .. For the expression of 10xHis-tagged GCase wt, a commercially available plasmid (Sino Biological Inc., Peking, China, #HG12038-CH) was modified to make the C-terminal His tag cleavable by tobacco etch virus (TEV) protease.

    Plasmid Preparation:

    Article Title: Functional Analyses of SATB2 Variants Reveal Pathogenicity Mechanisms Linked With SATB2-Associated Syndrome.
    Article Snippet: .. SATB2 proteins with the seven variants, including variants from previous reports with detailed descriptions of the oral phenotype (Kikuiri et al. 2018; Kurosaka et al. 2023; Li, Ye, and Su 2022), were constructed using expression with a wildtype (WT) SATB2 plasmid (Human SATB2 Gene ORF cDNA clone expression plasmid, N- Flag tag; Sino Biological Inc., Kanagawa, Japan) as the DNA template, using the QuickChange Lightening Site- directed Mutagenesis Kit (Agilent Technologies, La Jolla, CA, USA) according to the manufacturer's protocol. .. Variants were confirmed using the Applied Biosystems Big Dye Terminator Reaction kit and Applied Biosystems 3130xl Genetic Analyzer.

    FLAG-tag:

    Article Title: Functional Analyses of SATB2 Variants Reveal Pathogenicity Mechanisms Linked With SATB2-Associated Syndrome.
    Article Snippet: .. SATB2 proteins with the seven variants, including variants from previous reports with detailed descriptions of the oral phenotype (Kikuiri et al. 2018; Kurosaka et al. 2023; Li, Ye, and Su 2022), were constructed using expression with a wildtype (WT) SATB2 plasmid (Human SATB2 Gene ORF cDNA clone expression plasmid, N- Flag tag; Sino Biological Inc., Kanagawa, Japan) as the DNA template, using the QuickChange Lightening Site- directed Mutagenesis Kit (Agilent Technologies, La Jolla, CA, USA) according to the manufacturer's protocol. .. Variants were confirmed using the Applied Biosystems Big Dye Terminator Reaction kit and Applied Biosystems 3130xl Genetic Analyzer.

    Mutagenesis:

    Article Title: Functional Analyses of SATB2 Variants Reveal Pathogenicity Mechanisms Linked With SATB2-Associated Syndrome.
    Article Snippet: .. SATB2 proteins with the seven variants, including variants from previous reports with detailed descriptions of the oral phenotype (Kikuiri et al. 2018; Kurosaka et al. 2023; Li, Ye, and Su 2022), were constructed using expression with a wildtype (WT) SATB2 plasmid (Human SATB2 Gene ORF cDNA clone expression plasmid, N- Flag tag; Sino Biological Inc., Kanagawa, Japan) as the DNA template, using the QuickChange Lightening Site- directed Mutagenesis Kit (Agilent Technologies, La Jolla, CA, USA) according to the manufacturer's protocol. .. Variants were confirmed using the Applied Biosystems Big Dye Terminator Reaction kit and Applied Biosystems 3130xl Genetic Analyzer.



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    ( A ) Diagram of amino acid changes in <t>WT-MYRF,</t> dG-MYRF, and cleavage deficient variant V679A-MYRF. ProRich, proline rich; DBD, <t>DNA</t> binding domain; ICA, intramolecular chaperone auto-processing; TM, transmembrane; C2, C-terminal. ( B ) Localization <t>of</t> <t>FLAG-tagged</t> MYRF constructs in ARPE-19 cells show normal nuclear localization of dG-MYRF ( n = 3). Scale bar: 50 μm. ( C ) Western blot of transfected ARPE-19 cells shows no change in cleavage of N-terminal fragment ( n = 3). ( D ) qPCR analysis of RNA from ARPE-19 cells transduced with dG-MYRF, compared with WT-MYRF show decreased levels of transcripts for total Myr f and endogenous Tmem9 8 mRNA ( n = 3) by Student’s t test. * P < 0.05, ** P < 0.01.
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    ( A ) Diagram of amino acid changes in <t>WT-MYRF,</t> dG-MYRF, and cleavage deficient variant V679A-MYRF. ProRich, proline rich; DBD, <t>DNA</t> binding domain; ICA, intramolecular chaperone auto-processing; TM, transmembrane; C2, C-terminal. ( B ) Localization <t>of</t> <t>FLAG-tagged</t> MYRF constructs in ARPE-19 cells show normal nuclear localization of dG-MYRF ( n = 3). Scale bar: 50 μm. ( C ) Western blot of transfected ARPE-19 cells shows no change in cleavage of N-terminal fragment ( n = 3). ( D ) qPCR analysis of RNA from ARPE-19 cells transduced with dG-MYRF, compared with WT-MYRF show decreased levels of transcripts for total Myr f and endogenous Tmem9 8 mRNA ( n = 3) by Student’s t test. * P < 0.05, ** P < 0.01.
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    ( A ) Diagram of amino acid changes in <t>WT-MYRF,</t> dG-MYRF, and cleavage deficient variant V679A-MYRF. ProRich, proline rich; DBD, <t>DNA</t> binding domain; ICA, intramolecular chaperone auto-processing; TM, transmembrane; C2, C-terminal. ( B ) Localization <t>of</t> <t>FLAG-tagged</t> MYRF constructs in ARPE-19 cells show normal nuclear localization of dG-MYRF ( n = 3). Scale bar: 50 μm. ( C ) Western blot of transfected ARPE-19 cells shows no change in cleavage of N-terminal fragment ( n = 3). ( D ) qPCR analysis of RNA from ARPE-19 cells transduced with dG-MYRF, compared with WT-MYRF show decreased levels of transcripts for total Myr f and endogenous Tmem9 8 mRNA ( n = 3) by Student’s t test. * P < 0.05, ** P < 0.01.
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    ( A ) Diagram of amino acid changes in <t>WT-MYRF,</t> dG-MYRF, and cleavage deficient variant V679A-MYRF. ProRich, proline rich; DBD, <t>DNA</t> binding domain; ICA, intramolecular chaperone auto-processing; TM, transmembrane; C2, C-terminal. ( B ) Localization <t>of</t> <t>FLAG-tagged</t> MYRF constructs in ARPE-19 cells show normal nuclear localization of dG-MYRF ( n = 3). Scale bar: 50 μm. ( C ) Western blot of transfected ARPE-19 cells shows no change in cleavage of N-terminal fragment ( n = 3). ( D ) qPCR analysis of RNA from ARPE-19 cells transduced with dG-MYRF, compared with WT-MYRF show decreased levels of transcripts for total Myr f and endogenous Tmem9 8 mRNA ( n = 3) by Student’s t test. * P < 0.05, ** P < 0.01.
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    ( A ) Diagram of amino acid changes in <t>WT-MYRF,</t> dG-MYRF, and cleavage deficient variant V679A-MYRF. ProRich, proline rich; DBD, <t>DNA</t> binding domain; ICA, intramolecular chaperone auto-processing; TM, transmembrane; C2, C-terminal. ( B ) Localization <t>of</t> <t>FLAG-tagged</t> MYRF constructs in ARPE-19 cells show normal nuclear localization of dG-MYRF ( n = 3). Scale bar: 50 μm. ( C ) Western blot of transfected ARPE-19 cells shows no change in cleavage of N-terminal fragment ( n = 3). ( D ) qPCR analysis of RNA from ARPE-19 cells transduced with dG-MYRF, compared with WT-MYRF show decreased levels of transcripts for total Myr f and endogenous Tmem9 8 mRNA ( n = 3) by Student’s t test. * P < 0.05, ** P < 0.01.
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    Image Search Results


    ( A ) Diagram of amino acid changes in WT-MYRF, dG-MYRF, and cleavage deficient variant V679A-MYRF. ProRich, proline rich; DBD, DNA binding domain; ICA, intramolecular chaperone auto-processing; TM, transmembrane; C2, C-terminal. ( B ) Localization of FLAG-tagged MYRF constructs in ARPE-19 cells show normal nuclear localization of dG-MYRF ( n = 3). Scale bar: 50 μm. ( C ) Western blot of transfected ARPE-19 cells shows no change in cleavage of N-terminal fragment ( n = 3). ( D ) qPCR analysis of RNA from ARPE-19 cells transduced with dG-MYRF, compared with WT-MYRF show decreased levels of transcripts for total Myr f and endogenous Tmem9 8 mRNA ( n = 3) by Student’s t test. * P < 0.05, ** P < 0.01.

    Journal: JCI Insight

    Article Title: Splicing variants in MYRF cause partial loss of function in the retinal pigment epithelium leading to nanophthalmos

    doi: 10.1172/jci.insight.194681

    Figure Lengend Snippet: ( A ) Diagram of amino acid changes in WT-MYRF, dG-MYRF, and cleavage deficient variant V679A-MYRF. ProRich, proline rich; DBD, DNA binding domain; ICA, intramolecular chaperone auto-processing; TM, transmembrane; C2, C-terminal. ( B ) Localization of FLAG-tagged MYRF constructs in ARPE-19 cells show normal nuclear localization of dG-MYRF ( n = 3). Scale bar: 50 μm. ( C ) Western blot of transfected ARPE-19 cells shows no change in cleavage of N-terminal fragment ( n = 3). ( D ) qPCR analysis of RNA from ARPE-19 cells transduced with dG-MYRF, compared with WT-MYRF show decreased levels of transcripts for total Myr f and endogenous Tmem9 8 mRNA ( n = 3) by Student’s t test. * P < 0.05, ** P < 0.01.

    Article Snippet: Human FLAG-tagged WT-MYRF DNA constructs were generated (Twist Bioscience, South San Francisco, CA) in a pLentiLox-IRES-GFP backbone vector ( ).

    Techniques: Variant Assay, Binding Assay, Construct, Western Blot, Transfection, Transduction

    ( A – C ) Localization of FLAG-tagged MYRF constructs in mature human primary RPE show normal nuclear localization of dG-MYRF. Scale bar: 5 μm. ( D ) Western blot of transduced human primary RPE cells shows no change in cleavage of N-terminal fragment. ( E ) Functional assessment of barrier function via TEER (normalized to pretransduction TEER value for each replicate) shows no significant difference in TEER 1 week after transduction with Empty Vector, WT-MYRF, or dG-MYRF by 1-way ANOVA (all experiments are 3 reps from n = 2 donors).

    Journal: JCI Insight

    Article Title: Splicing variants in MYRF cause partial loss of function in the retinal pigment epithelium leading to nanophthalmos

    doi: 10.1172/jci.insight.194681

    Figure Lengend Snippet: ( A – C ) Localization of FLAG-tagged MYRF constructs in mature human primary RPE show normal nuclear localization of dG-MYRF. Scale bar: 5 μm. ( D ) Western blot of transduced human primary RPE cells shows no change in cleavage of N-terminal fragment. ( E ) Functional assessment of barrier function via TEER (normalized to pretransduction TEER value for each replicate) shows no significant difference in TEER 1 week after transduction with Empty Vector, WT-MYRF, or dG-MYRF by 1-way ANOVA (all experiments are 3 reps from n = 2 donors).

    Article Snippet: Human FLAG-tagged WT-MYRF DNA constructs were generated (Twist Bioscience, South San Francisco, CA) in a pLentiLox-IRES-GFP backbone vector ( ).

    Techniques: Construct, Western Blot, Functional Assay, Transduction, Plasmid Preparation

    WT MYRF retains transcription factor function resulting in no phenotypic effects. A C-terminal frameshift splice variant results in a 31 amino acid frameshift leading to decreased C-terminus stability and transcriptional activation, leading to reduced transcription factor function and isolated ocular disease. Intronic splicing variants lead to a frameshift that results in early truncation or nonsense mediated RNA decay. These splice sites are not always used, leading to modest decrease in MYRF transcription factor function and isolated ocular disease. In contrast, complete loss of function alleles either lead to early truncation/nonsense-mediated decay or alter critical functional domains (i.e., DNA binding, cleavage) and lead to severe, syndromic symptoms.

    Journal: JCI Insight

    Article Title: Splicing variants in MYRF cause partial loss of function in the retinal pigment epithelium leading to nanophthalmos

    doi: 10.1172/jci.insight.194681

    Figure Lengend Snippet: WT MYRF retains transcription factor function resulting in no phenotypic effects. A C-terminal frameshift splice variant results in a 31 amino acid frameshift leading to decreased C-terminus stability and transcriptional activation, leading to reduced transcription factor function and isolated ocular disease. Intronic splicing variants lead to a frameshift that results in early truncation or nonsense mediated RNA decay. These splice sites are not always used, leading to modest decrease in MYRF transcription factor function and isolated ocular disease. In contrast, complete loss of function alleles either lead to early truncation/nonsense-mediated decay or alter critical functional domains (i.e., DNA binding, cleavage) and lead to severe, syndromic symptoms.

    Article Snippet: Human FLAG-tagged WT-MYRF DNA constructs were generated (Twist Bioscience, South San Francisco, CA) in a pLentiLox-IRES-GFP backbone vector ( ).

    Techniques: Variant Assay, Activation Assay, Isolation, Functional Assay, Binding Assay