pcdna dest53  (Thermo Fisher)


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    Thermo Fisher pcdna dest53
    USP18 has no inhibitory effect on Jak/STAT signaling in Huh7.5 cells. Huh7.5 cells were seeded at 3 × 10 5 /ml, 2 ml per well in 6-well plates in antibiotic-free medium for 24 hours before 4 μ g blank vector <t>pcDNA-DEST53,</t> 4 μ g USP18 WT, or 4 μ g USP18 C64S was transfected into each well. 36 hours posttransfection, 100 IU/ml IFN α was added to each well. The cells were harvested at 0 min, 30 min, 2 hours, 4 hours, and 8 hours posttreatment. Total protein was extracted to detect USP18, phospho-STAT1 (Tyr701), and total STAT1 by western blot (a, upper), and phospho-STAT1 integrated density was normalized by STAT1 (a, bottom). Total RNA was extracted to detect ISG mRNAs by real-time PCR (b). V: transfected with 4 μ g blank vector pcDNA-DEST53; WT: transfected with 4 μ g USP18 WT; mu: transfected with 4 μ g USP18 C64S. Results are presented as means ± SD ( n ≥ 3).
    Pcdna Dest53, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pcdna dest53/product/Thermo Fisher
    Average 90 stars, based on 24 article reviews
    Price from $9.99 to $1999.99
    pcdna dest53 - by Bioz Stars, 2020-09
    90/100 stars

    Images

    1) Product Images from "The Ubiquitin-Specific Protease 18 Promotes Hepatitis C Virus Production by Increasing Viral Infectivity"

    Article Title: The Ubiquitin-Specific Protease 18 Promotes Hepatitis C Virus Production by Increasing Viral Infectivity

    Journal: Mediators of Inflammation

    doi: 10.1155/2019/3124745

    USP18 has no inhibitory effect on Jak/STAT signaling in Huh7.5 cells. Huh7.5 cells were seeded at 3 × 10 5 /ml, 2 ml per well in 6-well plates in antibiotic-free medium for 24 hours before 4 μ g blank vector pcDNA-DEST53, 4 μ g USP18 WT, or 4 μ g USP18 C64S was transfected into each well. 36 hours posttransfection, 100 IU/ml IFN α was added to each well. The cells were harvested at 0 min, 30 min, 2 hours, 4 hours, and 8 hours posttreatment. Total protein was extracted to detect USP18, phospho-STAT1 (Tyr701), and total STAT1 by western blot (a, upper), and phospho-STAT1 integrated density was normalized by STAT1 (a, bottom). Total RNA was extracted to detect ISG mRNAs by real-time PCR (b). V: transfected with 4 μ g blank vector pcDNA-DEST53; WT: transfected with 4 μ g USP18 WT; mu: transfected with 4 μ g USP18 C64S. Results are presented as means ± SD ( n ≥ 3).
    Figure Legend Snippet: USP18 has no inhibitory effect on Jak/STAT signaling in Huh7.5 cells. Huh7.5 cells were seeded at 3 × 10 5 /ml, 2 ml per well in 6-well plates in antibiotic-free medium for 24 hours before 4 μ g blank vector pcDNA-DEST53, 4 μ g USP18 WT, or 4 μ g USP18 C64S was transfected into each well. 36 hours posttransfection, 100 IU/ml IFN α was added to each well. The cells were harvested at 0 min, 30 min, 2 hours, 4 hours, and 8 hours posttreatment. Total protein was extracted to detect USP18, phospho-STAT1 (Tyr701), and total STAT1 by western blot (a, upper), and phospho-STAT1 integrated density was normalized by STAT1 (a, bottom). Total RNA was extracted to detect ISG mRNAs by real-time PCR (b). V: transfected with 4 μ g blank vector pcDNA-DEST53; WT: transfected with 4 μ g USP18 WT; mu: transfected with 4 μ g USP18 C64S. Results are presented as means ± SD ( n ≥ 3).

    Techniques Used: Plasmid Preparation, Transfection, Western Blot, Real-time Polymerase Chain Reaction

    USP18 overexpression does not affect HCV replicons nor microRNA122 expression. 5 × 10 5 AB12-A2 or sbJFH1-B2 cells were seeded overnight (6-well plates) before transfection with Lipofectamine 2000 in OptiMem with 4 μ g USP18 WT or USP18 C64S plasmid DNA. 24 hours posttransfection, protein was collected (RIPA buffer) or cells were treated with IFN α (Recombinant, Sigma) at 1, 100, 500, or 1000 IU/ml in DMEM without antibiotics and incubated for 72 h before RNA isolation. Total RNA was collected and subjected to real-time PCR assessment using primers specific for HCV Con1b (a) or HCV JFH1 5′ UTR (b). (c) Huh7.5 cells were transfected with 4 μ g blank vector pcDNA-DEST53, 4 μ g USP18 WT, or 4 μ g USP18 C64S for 48 hours before treatment with IFN α (10 IU/ml). MicroRNA 122 levels were determined by a kit and normalized to U6 at different time points indicated post-IFN α treatment. Results are presented as means ± SD ( n ≥ 3).
    Figure Legend Snippet: USP18 overexpression does not affect HCV replicons nor microRNA122 expression. 5 × 10 5 AB12-A2 or sbJFH1-B2 cells were seeded overnight (6-well plates) before transfection with Lipofectamine 2000 in OptiMem with 4 μ g USP18 WT or USP18 C64S plasmid DNA. 24 hours posttransfection, protein was collected (RIPA buffer) or cells were treated with IFN α (Recombinant, Sigma) at 1, 100, 500, or 1000 IU/ml in DMEM without antibiotics and incubated for 72 h before RNA isolation. Total RNA was collected and subjected to real-time PCR assessment using primers specific for HCV Con1b (a) or HCV JFH1 5′ UTR (b). (c) Huh7.5 cells were transfected with 4 μ g blank vector pcDNA-DEST53, 4 μ g USP18 WT, or 4 μ g USP18 C64S for 48 hours before treatment with IFN α (10 IU/ml). MicroRNA 122 levels were determined by a kit and normalized to U6 at different time points indicated post-IFN α treatment. Results are presented as means ± SD ( n ≥ 3).

    Techniques Used: Over Expression, Expressing, Transfection, Plasmid Preparation, Recombinant, Incubation, Isolation, Real-time Polymerase Chain Reaction

    Human USP18 expression and catalytic activity in Huh7.5 cells. (a) Huh7.5 cells were seeded at 3 × 10 5 /ml, 2 ml per well in 6-well plates in antibiotic-free medium for 24 hours before 4 μ g empty vector pcDNA-DEST53, 2 μ g USP18 WT, or 4 μ g USP18 WT was transfected into each well. 48 hours posttransfection, total protein was extracted to detect GFP tag by western blot. (b) Cleavage of ISG15-GST fusion in vitro. Huh7.5 cells were transfected with various USP18 plasmids (wild-type USP18, WT; USP18 C64S, C64S; USP18 C65S, C65S; or USP18 C64/65S, C64/65S) in combination with ISG15-GST. 48 hours posttransfection, total protein was extracted to detect ISG15 and USP18 by western blot. (c) Cleavage of ISG15 conjugates in IFN α -treated Huh7.5 cells. Huh7.5 cells were transfected with an empty vector (vector), USP18 WT, or USP18 C64S. 24 hrs later, the cells were treated with IFN α (0-100 U/ml) for 24 hours, after which western bot was performed to analyze expressions of ISG15 conjugates and USP18. Untreated: untreated control; mock: transfected with 4 μ g empty vector pcDNA-DEST53; pcDNA-DEST53-USP18: transfected with wild-type USP18 (USP18 WT).
    Figure Legend Snippet: Human USP18 expression and catalytic activity in Huh7.5 cells. (a) Huh7.5 cells were seeded at 3 × 10 5 /ml, 2 ml per well in 6-well plates in antibiotic-free medium for 24 hours before 4 μ g empty vector pcDNA-DEST53, 2 μ g USP18 WT, or 4 μ g USP18 WT was transfected into each well. 48 hours posttransfection, total protein was extracted to detect GFP tag by western blot. (b) Cleavage of ISG15-GST fusion in vitro. Huh7.5 cells were transfected with various USP18 plasmids (wild-type USP18, WT; USP18 C64S, C64S; USP18 C65S, C65S; or USP18 C64/65S, C64/65S) in combination with ISG15-GST. 48 hours posttransfection, total protein was extracted to detect ISG15 and USP18 by western blot. (c) Cleavage of ISG15 conjugates in IFN α -treated Huh7.5 cells. Huh7.5 cells were transfected with an empty vector (vector), USP18 WT, or USP18 C64S. 24 hrs later, the cells were treated with IFN α (0-100 U/ml) for 24 hours, after which western bot was performed to analyze expressions of ISG15 conjugates and USP18. Untreated: untreated control; mock: transfected with 4 μ g empty vector pcDNA-DEST53; pcDNA-DEST53-USP18: transfected with wild-type USP18 (USP18 WT).

    Techniques Used: Expressing, Activity Assay, Plasmid Preparation, Transfection, Western Blot, In Vitro

    Protease-independent promotion of HCV production and blunting of type I and type III IFN anti-HCV activity by USP18. Huh7.5 cells were seeded at 1.5 × 10 5 /well in 12-well plates one day before 2 μ g blank vector pcDNA-DEST53, 2 μ g USP18 WT, or 2 μ g USP18 C64S was transfected. 48 hours posttransfection, J6/JFH1 virus was added (MOI = 4) and incubated for 4 hours before the culture medium was removed. And then, the cells were washed and supplied with fresh medium and cultured for another 48 hours. The intracellular total RNA and the culture medium were collected. J6/JFH1 RNA (a) and J6/JFH1 virion production (b, c) in the presence and absence of IFN α or IFN λ were detected by real-time PCR or limiting dilution analysis, respectively. Results are presented as means ± SD ( n ≥ 3). ∗ p
    Figure Legend Snippet: Protease-independent promotion of HCV production and blunting of type I and type III IFN anti-HCV activity by USP18. Huh7.5 cells were seeded at 1.5 × 10 5 /well in 12-well plates one day before 2 μ g blank vector pcDNA-DEST53, 2 μ g USP18 WT, or 2 μ g USP18 C64S was transfected. 48 hours posttransfection, J6/JFH1 virus was added (MOI = 4) and incubated for 4 hours before the culture medium was removed. And then, the cells were washed and supplied with fresh medium and cultured for another 48 hours. The intracellular total RNA and the culture medium were collected. J6/JFH1 RNA (a) and J6/JFH1 virion production (b, c) in the presence and absence of IFN α or IFN λ were detected by real-time PCR or limiting dilution analysis, respectively. Results are presented as means ± SD ( n ≥ 3). ∗ p

    Techniques Used: Activity Assay, Plasmid Preparation, Transfection, Incubation, Cell Culture, Real-time Polymerase Chain Reaction

    Related Articles

    Clone Assay:

    Article Title: Perturbation of PALB2 function by the T413S mutation found in small cell lung cancer
    Article Snippet: .. For GFP-ChAM expression in HEK293T cells, ChAM variants in pENTR1A were transferred to pcDNA-DEST53 (12288015, Invitrogen) using Gateway cloning. .. For FLAG-PALB2 expression in EUFA1341 cells, PALB2 variants in pENTR3C were transferred to pCEP4-GW/N3xFLAG using Gateway cloning.

    Article Title: SPATA2 links CYLD to the TNF‐α receptor signaling complex and modulates the receptor signaling outcomes
    Article Snippet: .. Full‐length SPATA2 and CYLD were obtained from the human Ultimate ORFeome clones collection (Thermo Fisher Scientific) and cloned into Gateway‐compatible expression vectors pcDNA‐DEST53 (Thermo Fisher Scientific) and pMX‐DEST53‐TwinStrep‐FLAG (custom made) for N‐terminal tagging of SPATA2 with GFP and CYLD with TwinStrep‐FLAG. .. N‐terminally tagged GFP‐SPATA2 458* (aa1–458), 200* (aa1–200) and D200 (aa221–520) and Strep‐FLAG CYLD 600* (aa1–600), 400* (aa1–400) and USP (aa592–950) mutants were generated by site‐directed mutagenesis.

    Article Title: Nol9 is a novel polynucleotide 5?-kinase involved in ribosomal RNA processing
    Article Snippet: .. In the second round of transfection with the pSUPER vector, we co-transfected wild type or mutant Nol9 cloned into the pcDNA-DEST53 Gateway vector (Invitrogen). .. Metabolic labelling Cells were labelled with L -[methyl-3 H]-methionine as described in ).

    Article Title: ATG9A shapes the forming autophagosome through Arfaptin 2 and phosphatidylinositol 4-kinase IIIβ
    Article Snippet: .. Expression clones were made as described in the Gateway cloning technology instruction manual (Invitrogen) using the gateway expression vectors pDestHA ( ) and pDest53 (Invitrogen; 12288015). pDestHA-ARFIP2 W99A point mutation was generated by using QuikChange Multi Site-Directed Mutagenesis Kit (Agilent Technologies; 200515). iRFP-PI4KIIIβ was generated by subcloning PI4KIIIβ into pLVX-iRFP. ..

    Transfection:

    Article Title: ATG4B contains a C-terminal LIR motif important for binding and efficient cleavage of mammalian orthologs of yeast Atg8
    Article Snippet: .. Gateway destination vectors used were pDEST15 (bacterial expression of N-terminal GST fusion proteins; Thermo Fisher Scientific, 11802014), pDEST24 (bacterial expression of C-terminal GST fusion proteins) (Thermo Fisher Scientific, 12216016), pDESTmyc (mammalian expression of N-terminal MYC-tagged proteins), pDEST-EGFP (mammalian expression of N-terminal EGFP-tagged proteins), and pDEST53 (mammalian expression of N-terminal GFP-tagged proteins; Thermo Fisher Scientific, 12288015). pDEST-LTR-EGFP (mammalian transfection vector for stable and doxycycline controlled inducible expression of N-terminal-EGFP tagged fusion constructs under the control of a truncated CMV promoter) conferring blasticidin resistance was made by subcloning of EGFP and the GATEWAY cassette as a PCR product from pDEST-EGFP-C1 into the reverse Tetracycline retrovirus vector pLRT-X (a kind gift from Dr. Masaaki Komatsu, School of Medicine, Niigata University, Japan). .. Transfer of pENTR constructs to destination vectors was performed using the Gateway LR reaction (Thermo Fisher Scientific, 11791020).

    Article Title: Nol9 is a novel polynucleotide 5?-kinase involved in ribosomal RNA processing
    Article Snippet: .. In the second round of transfection with the pSUPER vector, we co-transfected wild type or mutant Nol9 cloned into the pcDNA-DEST53 Gateway vector (Invitrogen). .. Metabolic labelling Cells were labelled with L -[methyl-3 H]-methionine as described in ).

    Mutagenesis:

    Article Title: Nol9 is a novel polynucleotide 5?-kinase involved in ribosomal RNA processing
    Article Snippet: .. In the second round of transfection with the pSUPER vector, we co-transfected wild type or mutant Nol9 cloned into the pcDNA-DEST53 Gateway vector (Invitrogen). .. Metabolic labelling Cells were labelled with L -[methyl-3 H]-methionine as described in ).

    Article Title: ATG9A shapes the forming autophagosome through Arfaptin 2 and phosphatidylinositol 4-kinase IIIβ
    Article Snippet: .. Expression clones were made as described in the Gateway cloning technology instruction manual (Invitrogen) using the gateway expression vectors pDestHA ( ) and pDest53 (Invitrogen; 12288015). pDestHA-ARFIP2 W99A point mutation was generated by using QuikChange Multi Site-Directed Mutagenesis Kit (Agilent Technologies; 200515). iRFP-PI4KIIIβ was generated by subcloning PI4KIIIβ into pLVX-iRFP. ..

    Subcloning:

    Article Title: ATG4B contains a C-terminal LIR motif important for binding and efficient cleavage of mammalian orthologs of yeast Atg8
    Article Snippet: .. Gateway destination vectors used were pDEST15 (bacterial expression of N-terminal GST fusion proteins; Thermo Fisher Scientific, 11802014), pDEST24 (bacterial expression of C-terminal GST fusion proteins) (Thermo Fisher Scientific, 12216016), pDESTmyc (mammalian expression of N-terminal MYC-tagged proteins), pDEST-EGFP (mammalian expression of N-terminal EGFP-tagged proteins), and pDEST53 (mammalian expression of N-terminal GFP-tagged proteins; Thermo Fisher Scientific, 12288015). pDEST-LTR-EGFP (mammalian transfection vector for stable and doxycycline controlled inducible expression of N-terminal-EGFP tagged fusion constructs under the control of a truncated CMV promoter) conferring blasticidin resistance was made by subcloning of EGFP and the GATEWAY cassette as a PCR product from pDEST-EGFP-C1 into the reverse Tetracycline retrovirus vector pLRT-X (a kind gift from Dr. Masaaki Komatsu, School of Medicine, Niigata University, Japan). .. Transfer of pENTR constructs to destination vectors was performed using the Gateway LR reaction (Thermo Fisher Scientific, 11791020).

    Article Title: ATG9A shapes the forming autophagosome through Arfaptin 2 and phosphatidylinositol 4-kinase IIIβ
    Article Snippet: .. Expression clones were made as described in the Gateway cloning technology instruction manual (Invitrogen) using the gateway expression vectors pDestHA ( ) and pDest53 (Invitrogen; 12288015). pDestHA-ARFIP2 W99A point mutation was generated by using QuikChange Multi Site-Directed Mutagenesis Kit (Agilent Technologies; 200515). iRFP-PI4KIIIβ was generated by subcloning PI4KIIIβ into pLVX-iRFP. ..

    Construct:

    Article Title: ATG4B contains a C-terminal LIR motif important for binding and efficient cleavage of mammalian orthologs of yeast Atg8
    Article Snippet: .. Gateway destination vectors used were pDEST15 (bacterial expression of N-terminal GST fusion proteins; Thermo Fisher Scientific, 11802014), pDEST24 (bacterial expression of C-terminal GST fusion proteins) (Thermo Fisher Scientific, 12216016), pDESTmyc (mammalian expression of N-terminal MYC-tagged proteins), pDEST-EGFP (mammalian expression of N-terminal EGFP-tagged proteins), and pDEST53 (mammalian expression of N-terminal GFP-tagged proteins; Thermo Fisher Scientific, 12288015). pDEST-LTR-EGFP (mammalian transfection vector for stable and doxycycline controlled inducible expression of N-terminal-EGFP tagged fusion constructs under the control of a truncated CMV promoter) conferring blasticidin resistance was made by subcloning of EGFP and the GATEWAY cassette as a PCR product from pDEST-EGFP-C1 into the reverse Tetracycline retrovirus vector pLRT-X (a kind gift from Dr. Masaaki Komatsu, School of Medicine, Niigata University, Japan). .. Transfer of pENTR constructs to destination vectors was performed using the Gateway LR reaction (Thermo Fisher Scientific, 11791020).

    FLAG-tag:

    Article Title: EVI1 oncoprotein interacts with a large and complex network of proteins and integrates signals through protein phosphorylation
    Article Snippet: .. EVI1 cDNA was transferred into a Gateway pcDNA-DEST53 vector (Invitrogen) for which the GFP tag had been replaced with a FLAG tag (N terminus). .. Mutations in the FLAG-EVI1 plasmid were generated by site-directed mutagenesis.

    Generated:

    Article Title: ATG9A shapes the forming autophagosome through Arfaptin 2 and phosphatidylinositol 4-kinase IIIβ
    Article Snippet: .. Expression clones were made as described in the Gateway cloning technology instruction manual (Invitrogen) using the gateway expression vectors pDestHA ( ) and pDest53 (Invitrogen; 12288015). pDestHA-ARFIP2 W99A point mutation was generated by using QuikChange Multi Site-Directed Mutagenesis Kit (Agilent Technologies; 200515). iRFP-PI4KIIIβ was generated by subcloning PI4KIIIβ into pLVX-iRFP. ..

    Expressing:

    Article Title: Identification and Characterisation of the RalA-ERp57 Interaction: Evidence for GDI Activity of ERp57
    Article Snippet: .. The ORFs were further sub-cloned by recombination into pDEST15 (bacterial N-terminal GST-tag), pDEST17 (bacterial N-terminal 6xHis-tag) and pcDNA-DEST53 (mammalian N-terminal GFP-tag) expression vectors (Invitrogen). .. Expression of 6xHis-ERp57 in Escherichia coli and purification using Ni-NTA resin (Qiagen) were carried out according to the manufacturer’s protocol.

    Article Title: Perturbation of PALB2 function by the T413S mutation found in small cell lung cancer
    Article Snippet: .. For GFP-ChAM expression in HEK293T cells, ChAM variants in pENTR1A were transferred to pcDNA-DEST53 (12288015, Invitrogen) using Gateway cloning. .. For FLAG-PALB2 expression in EUFA1341 cells, PALB2 variants in pENTR3C were transferred to pCEP4-GW/N3xFLAG using Gateway cloning.

    Article Title: ATG4B contains a C-terminal LIR motif important for binding and efficient cleavage of mammalian orthologs of yeast Atg8
    Article Snippet: .. Gateway destination vectors used were pDEST15 (bacterial expression of N-terminal GST fusion proteins; Thermo Fisher Scientific, 11802014), pDEST24 (bacterial expression of C-terminal GST fusion proteins) (Thermo Fisher Scientific, 12216016), pDESTmyc (mammalian expression of N-terminal MYC-tagged proteins), pDEST-EGFP (mammalian expression of N-terminal EGFP-tagged proteins), and pDEST53 (mammalian expression of N-terminal GFP-tagged proteins; Thermo Fisher Scientific, 12288015). pDEST-LTR-EGFP (mammalian transfection vector for stable and doxycycline controlled inducible expression of N-terminal-EGFP tagged fusion constructs under the control of a truncated CMV promoter) conferring blasticidin resistance was made by subcloning of EGFP and the GATEWAY cassette as a PCR product from pDEST-EGFP-C1 into the reverse Tetracycline retrovirus vector pLRT-X (a kind gift from Dr. Masaaki Komatsu, School of Medicine, Niigata University, Japan). .. Transfer of pENTR constructs to destination vectors was performed using the Gateway LR reaction (Thermo Fisher Scientific, 11791020).

    Article Title: SPATA2 links CYLD to the TNF‐α receptor signaling complex and modulates the receptor signaling outcomes
    Article Snippet: .. Full‐length SPATA2 and CYLD were obtained from the human Ultimate ORFeome clones collection (Thermo Fisher Scientific) and cloned into Gateway‐compatible expression vectors pcDNA‐DEST53 (Thermo Fisher Scientific) and pMX‐DEST53‐TwinStrep‐FLAG (custom made) for N‐terminal tagging of SPATA2 with GFP and CYLD with TwinStrep‐FLAG. .. N‐terminally tagged GFP‐SPATA2 458* (aa1–458), 200* (aa1–200) and D200 (aa221–520) and Strep‐FLAG CYLD 600* (aa1–600), 400* (aa1–400) and USP (aa592–950) mutants were generated by site‐directed mutagenesis.

    Article Title: ATG9A shapes the forming autophagosome through Arfaptin 2 and phosphatidylinositol 4-kinase IIIβ
    Article Snippet: .. Expression clones were made as described in the Gateway cloning technology instruction manual (Invitrogen) using the gateway expression vectors pDestHA ( ) and pDest53 (Invitrogen; 12288015). pDestHA-ARFIP2 W99A point mutation was generated by using QuikChange Multi Site-Directed Mutagenesis Kit (Agilent Technologies; 200515). iRFP-PI4KIIIβ was generated by subcloning PI4KIIIβ into pLVX-iRFP. ..

    Polymerase Chain Reaction:

    Article Title: ATG4B contains a C-terminal LIR motif important for binding and efficient cleavage of mammalian orthologs of yeast Atg8
    Article Snippet: .. Gateway destination vectors used were pDEST15 (bacterial expression of N-terminal GST fusion proteins; Thermo Fisher Scientific, 11802014), pDEST24 (bacterial expression of C-terminal GST fusion proteins) (Thermo Fisher Scientific, 12216016), pDESTmyc (mammalian expression of N-terminal MYC-tagged proteins), pDEST-EGFP (mammalian expression of N-terminal EGFP-tagged proteins), and pDEST53 (mammalian expression of N-terminal GFP-tagged proteins; Thermo Fisher Scientific, 12288015). pDEST-LTR-EGFP (mammalian transfection vector for stable and doxycycline controlled inducible expression of N-terminal-EGFP tagged fusion constructs under the control of a truncated CMV promoter) conferring blasticidin resistance was made by subcloning of EGFP and the GATEWAY cassette as a PCR product from pDEST-EGFP-C1 into the reverse Tetracycline retrovirus vector pLRT-X (a kind gift from Dr. Masaaki Komatsu, School of Medicine, Niigata University, Japan). .. Transfer of pENTR constructs to destination vectors was performed using the Gateway LR reaction (Thermo Fisher Scientific, 11791020).

    Plasmid Preparation:

    Article Title: Fabrication of Functionalized Double-Lamellar Multifunctional Envelope-Type Nanodevices Using a Microfluidic Chip with a Chaotic Mixer Array
    Article Snippet: .. HEPES buffer solution (10 mM, pH 7.4) was purchase from Dojindo (Japan). pcDNA-DEST53 Gateway vector was acquired from Invitrogen (USA). .. STR-R8 was obtained from Kurabo Industries (Japan).

    Article Title: ATG4B contains a C-terminal LIR motif important for binding and efficient cleavage of mammalian orthologs of yeast Atg8
    Article Snippet: .. Gateway destination vectors used were pDEST15 (bacterial expression of N-terminal GST fusion proteins; Thermo Fisher Scientific, 11802014), pDEST24 (bacterial expression of C-terminal GST fusion proteins) (Thermo Fisher Scientific, 12216016), pDESTmyc (mammalian expression of N-terminal MYC-tagged proteins), pDEST-EGFP (mammalian expression of N-terminal EGFP-tagged proteins), and pDEST53 (mammalian expression of N-terminal GFP-tagged proteins; Thermo Fisher Scientific, 12288015). pDEST-LTR-EGFP (mammalian transfection vector for stable and doxycycline controlled inducible expression of N-terminal-EGFP tagged fusion constructs under the control of a truncated CMV promoter) conferring blasticidin resistance was made by subcloning of EGFP and the GATEWAY cassette as a PCR product from pDEST-EGFP-C1 into the reverse Tetracycline retrovirus vector pLRT-X (a kind gift from Dr. Masaaki Komatsu, School of Medicine, Niigata University, Japan). .. Transfer of pENTR constructs to destination vectors was performed using the Gateway LR reaction (Thermo Fisher Scientific, 11791020).

    Article Title: Nol9 is a novel polynucleotide 5?-kinase involved in ribosomal RNA processing
    Article Snippet: .. In the second round of transfection with the pSUPER vector, we co-transfected wild type or mutant Nol9 cloned into the pcDNA-DEST53 Gateway vector (Invitrogen). .. Metabolic labelling Cells were labelled with L -[methyl-3 H]-methionine as described in ).

    Article Title: EVI1 oncoprotein interacts with a large and complex network of proteins and integrates signals through protein phosphorylation
    Article Snippet: .. EVI1 cDNA was transferred into a Gateway pcDNA-DEST53 vector (Invitrogen) for which the GFP tag had been replaced with a FLAG tag (N terminus). .. Mutations in the FLAG-EVI1 plasmid were generated by site-directed mutagenesis.

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    Thermo Fisher gateway pcdna dest53 vector
    Gateway Pcdna Dest53 Vector, supplied by Thermo Fisher, 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/result/gateway pcdna dest53 vector/product/Thermo Fisher
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    Thermo Fisher pcdna dest53 vector
    Pcdna Dest53 Vector, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 42 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pcdna dest53 vector/product/Thermo Fisher
    Average 90 stars, based on 42 article reviews
    Price from $9.99 to $1999.99
    pcdna dest53 vector - by Bioz Stars, 2020-09
    90/100 stars
      Buy from Supplier

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