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Pcdna3.1d v5 his vector

Manufactured by Thermo Fisher Scientific
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The PcDNA3.1D/V5-His vector is a plasmid used for the expression of recombinant proteins in mammalian cell lines. It contains a cytomegalovirus (CMV) promoter for high-level expression, a multiple cloning site for insertion of the gene of interest, and a C-terminal V5 epitope tag and polyhistidine (6xHis) tag for detection and purification of the expressed protein.

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Lab products found in correlation

Quikchange 2 xl site directed mutagenesis kit, by Agilent Technologies (2 mentions) X tremegene hp dna transfection reagent, by Roche (2 mentions) Phusion high fidelity dna polymerase kit, by New England Biolabs (2 mentions) Turbofect transfection reagent, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

PcDNA3.1 (2401 citations) PcDNA3 vector (212 citations) PcDNA3.1D/V5-His-TOPO vector (21 citations) PcDNA3.1D (6 citations) PcDNA3.1D/v5-his (5 citations) PcDNA3.1D/V5-His-TOPO expression vector (5 citations)

4 protocols using pcdna3.1d v5 his vector

1

Molecular Cloning and Expression of Ubiquitin-Related Proteins

Cited 2 times
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The cDNA encoding HA-ubiquitin was a gift from Dr Yutong Zhao. NALP7, STAMBP, STAMBPL1, USP7, COPS5 and USP8 cDNA with complete protein coding gene sequences, all in pLX304 vector, were purchased from DNASU. Flag-HA-BRCC3 (Addgene plasmid #22540) was a gift from Wade Harper50 (link). pFBD-ESCRT 0 (encoding STAM and HRS) was a gift from James Hurley (Addgene plasmid # 21499)51 (link). The coding sequences from NALP7, STAM, STAMBP and BRCC3 plasmids as well as the NALP7 truncation mutants were cloned using the Phusion High-Fidelity DNA Polymerase kit (New England BioLabs). Site-directed mutagenesis was performed using the QuikChange II XL Site-Directed Mutagenesis kit (Agilent). All primers were purchased from Integrated DNA Technologies (IDT), and sequences are available in Supplementary Table 1. After cloning, cDNA was ligated into pcDNA3.1D/V5-His vector (Invitrogen) and constructs were verified by Sanger sequencing. Plasmids were overexpressed in HeLa cells using Turbofect transfection reagent (Thermo Scientific) or Beas2B cells using XtremeGene HP DNA Transfection Reagent (Roche) for 24–48 h before assay.
Bednash J.S., Weathington N., Londino J., Rojas M., Gulick D.L., Fort R., Han S., McKelvey A.C., Chen B.B, & Mallampalli R.K. (2017). Targeting the deubiquitinase STAMBP inhibits NALP7 inflammasome activity. Nature Communications, 8, 15203.
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2

Site-Directed Mutagenesis of TTF1 Lysine

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TTF1, HECW1, NEDD4L and hWWP1 cDNA were inserted into a pcDNA3.1D/V5-His vector or pcDNA3.1D/HA-His vector (Invitrogen, CA, USA) respectively. Site directed mutagenesis was performed to generate TTF1 lysine mutant according to the manufacturer’s instructions (Agilent Technologies, Santa Clara, CA, USA).
Liu J., Dong S., Li L., Wang H., Zhao J, & Zhao Y. (2019). The E3 ubiquitin ligase HECW1 targets thyroid transcription factor 1 (TTF1/NKX2.1) for its degradation in the ubiquitin-proteasome system. Cellular signalling, 58, 91-98.
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3

Overexpression of NLRP3, STAMBP, and USP8

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NLRP3, STAMBP, and USP8 cDNA with complete protein coding gene sequences, all in pLX304 vector, were purchased from DNASU. The coding sequences from NLRP3, STAMBP and USP8 plasmids were cloned using the Phusion High-Fidelity DNA Polymerase kit (New England BioLabs). Site-directed mutagenesis was performed using the QuikChange II XL Site-Directed Mutagenesis kit (Agilent). After cloning, cDNA was ligated into pcDNA3.1D/V5-His vector (Invitrogen) and constructs were verified by Sanger sequencing. Plasmids were overexpressed in HEK-293 cells using XtremeGene HP DNA Transfection Reagent (Roche) for 24–48 h before assay.
Bednash J.S., Johns F., Patel N., Smail T.R., Londino J.D, & Mallampalli R.K. (2020). The deubiquitinase STAMBP modulates cytokine secretion through the NLRP3 inflammasome. Cellular signalling, 79, 109859.
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4

Rac3 Mutant Generation via FBXL19 cDNA

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The FBXL19 cDNA was inserted into a pcDNA3.1D/V5-His vector (Invitrogen, CA, USA). Site directed mutagenesis was performed to generate Rac3 lysine or serine mutant according to the manufacturer’s instructions (Agilent Technologies, Santa Clara, CA, USA).
Dong S., Zhao J., Wei J., Bowser R.K., Khoo A., Liu Z., Luketich J.D., Pennathur A., Ma H, & Zhao Y. (2014). F-box protein complex FBXL19 regulates TGFβ1-induced E-cadherin down-regulation by mediating Rac3 ubiquitination and degradation. Molecular Cancer, 13, 76.
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