2024 Construction of pcdna3.i

Construction of pcdna3.i

Author: ooyx

August undefined, 2024

WebMar 14, 2024 · We provide a detailed, step-by-step protocol for donor vector design and construction using the pcDNA3 vector . Custom donor vectors can be difficult to clone and expensive to purchase. We provide a simple, efficient protocol to obtain custom donor vectors from the common pcDNA3 mammalian expression vector. WebDec 31, 2024 · Furthermore, pcDNA3 expressing EGFP was used as a control for background level signaling and for evaluating transfection efficiency, except for bioplex experiments, for which pcDNA3 alone was used. The integrity of all constructs was confirmed by sequencing. The extracellular chemokine domain of human fractalkine was …

Construction and development of a mammalian cell …

WebApr 10, 2024 · The DUBs (including ATXN3, BRCC3, COPS5, USP15, USP47, UCHL1, OTUB1, OTUD6B, and VCPIP1) were cloned into the pcDNA3 vector. The OTUD6B shRNA, β-TrCP, and SNAIL shRNA sequences were cloned into the pSIH-H1 vector. The siRNAs targeting RARα were purchased from RiboBio (Guangzhou, China). WebNov 8, 2024 · ( A) The pcDNA3.1 (+)/MgPa-transfected SV-HUC-1 cells were detected using indirect immunofluorescence (100×). SV-HUC-1 cells were fixed, blocked, and incubated with anti-MgPa antibody followed by incubation with Cy3-conjugated AffiniPure goat anti-rabbit IgG (H + G), then the cells were incubated with DAPI staining solution … baranaismartiktok

A protocol for custom CRISPR Cas9 donor vector construction to …

WebOct 26, 2024 · The pcDNA3.1 vector is a plasmid commonly used in recombinant protein expression in mammalian cells. This plasmid is also widely used to develop recombinant DNA-based vaccines for infectious diseases and other purposes, such as cancer gene therapy [ 15, 16, 17, 18 ]. WebOct 19, 2024 · The following vectors were used for construction: pcDNA3.1(+) (Invitrogen, United States), pGL4.10[luc2] (Promega), and pEGFP-C3 (Clontech, United States). The pRL-tk (Promega) plasmid was used to normalize luciferase activity. Plasmid DNA was purified with a Plasmid Miniprep kit. The DNA fragments were isolated from agarose gel … baranaika

Construction of targeted plasmid vector pcDNA3.1-Egr.1p …

[Construction of bicistronic vector and its application to

WebpcDNA3.1-e green fluorescent protein (GFP) is an important compound that is already established and widely used as a marker in biomolecular works. Producing pcDNA3.1-eGFP is not very complicated. ... Meshkat Z. Designing and construction of PCDNA 3.1 vector encoding CFP10 the in-vivo transfection of pcDNA3.1 led to significant inhibition gene ... WebApr 7, 2024 · Plasmid and Cell Line Construction. pCDNA3-GFP-(CUG) 5 and pCDNA3-GFP-(CUG) 200 plasmids were as previously reported . In such plasmids, 5 and 200 CTG repeats were placed at the 3’ UTR regions of the GFP gene, respectively. In this study, the pLL4.0 backbone was generated by replacing a GFP expressing cassette in the pLL3.7 … baranagar temperatureWebThe pcDNA3.1+/mtb32C plasmid was transformed into E. coli JM109 and then extracted. The mpt51 gene and pcDNA3.1+/mtb32C plasmid were both digested with EcoRI and BamHI restriction enzymes... baranailt

"WebGene amplification and construction of the recombinant plasmids The tPA-SP (GenBank accession no. E02360), MTQ, and MTI genes were synthesized by the Sangon Company (Shanghai, China) and then inserted into pcDNA3.1, as described above. The plasmids containing either 22P/A or 22P/G were constructed via site-directed mutagenesis. " - Construction of pcdna3.i

Construction of pcdna3.i

[A study on construction of plasmid pcDNA3.1 (-) CMV.CD and ...

WebpcDNA3.1 (-) CMV. CD was successfully constructed and CD-expressing Hep-2 cells can be killed by 5-FC, so the recombinant plasmid may be a candidate vector for laryngeal cancer therapy. [A study on construction of plasmid pcDNA3.1 (-) CMV.CD and transfection into laryngeal cancer cell Hep-2] Lin Chuang Er Bi Yan Hou Ke Za Zhi. WebJul 14, 2024 · pcDNA3.1 (+) and pcDNA3.1 (-) are 5.4 kb vectors derived from pcDNA3 and designed for high-level stable and transient expression in mammalian hosts. The high-level stable and non-replicative transient expression can be carried out in …

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Webconstruction pcDNA3.1 vector and the PCR product of IL-33 were double digested by EcoRI and NotI and Figure 2. IL-33 and Fra-1 protein expression after pcDNA-IL33 transfection. *P < 0.05, compared with control. Figure 3. Transwell boyden chamber detection of IL-33 impact on cell inva-sion. *P < 0.05, compared with control. BioRad … WebMar 14, 2024 · We provide a detailed protocol on how to design and construct a custom donor vector using Gibson assembly (and the commonly utilized pcDNA3 vector as the backbone) allowing researchers to obtain specific gene modifications of interest (gene truncation, gene deletion, epitope tagging or knock-in mutation).

WebHence, a DNA vaccine, named pCDNA3.1 (+)-MCP-Flag, was constructed by inserting the cloned LMBV major capsid protein (MCP) gene into the pCDNA3.1 (+)-Flag plasmid. The expression of the recombinant plasmid was confirmed by Western blot (WB) and RT-PCR. WebTo construct pcDNA3.1 (+)/A2E eukaryotic expression vector and obtain a stable expression on HLA-I negative human K562 cell, PCR technique was employed to amplify A2E cDNA from the multi-cistron expression vector pG/A2E carrying HLA-E and HLA-A2 cDNA through internal ribozyme entry site (IRES), the cDNA was subcloned into vector …

WebApr 10, 2024 · In addition, the pcDNA3.1(+)/CPSIT_p7 vaccine diminished pulmonary pathological lesions and reduced the C. psittaci load in the lungs of infected mice. It is worth noting that pcDNA3.1(+)/CPSIT_p7 suppressed C. psittaci dissemination in BALB/c mice. ... Briefly, the construction method of pcDNA3.1(+)/CPSIT_p7 plasmid was that the target … WebOct 26, 2015 · Mycobacterium tuberculosis is an etiological agent of human tuberculosis (TB). Designing new vaccines, including DNA vaccines, may be a useful strategy for …

WebContents: First plasmid construction：pCDNA3.0-PGK-BSD Annotation：pCDNA3.0 is a backbone plasmid ideal for transient expression in our designated cell lines including HEK293T,HepG2 and Bocs. All elements of our project are first to be incised into pCDNA3.0 and tested during transient expression in cell lines.

WebMar 29, 2006 · Construction of pcDNA3.1-based vectors with blasticidin and puromycin resistance markers. Masahiro Asada Signaling Molecules Research Laboratory, National … baranambalajWebpcDNA3 Mammalian expression vector with the CMV promoter and a neomycin-resistance marker. Sequence Author: Thermo Fisher (Invitrogen) Open in SnapGene Try SnapGene for Free Download Plasmid Download SnapGene Viewer Explore Over 2.7kPlasmids:Basic Cloning Vectors More Plasmid Sets No matches HomePlasmidsBasic Cloning … baranagar to sealdah train time tableWebWe present a detailed method for constructing a mammalian cell-based full-length antibody display library for targeting hepatocellular carcinoma. Two novel mammalian library vectors pcDNA3-CHm and pcDNA3-CLm were constructed that contained restriction enzyme sites NheI, ClaI and antibody constant domain. baranaismar3WebUniversity of Ostrava I guess CMV promoter (a strong promoter) + the enhanced stability of mRNA for the presence of SV40 polyA signal both part of the pcDNA3.1 plasmid will help strongly the... baranandishehWebOct 26, 2015 · After digesting the PCR product and the plasmid, the cfp10 fragment was ligated into the vector pcDNA3.1 (+). Correct insertion was confirmed by colony PCR, restriction enzyme digestion, and sequencing. Results: Electrophoresis of the PCR product on gel showed a 303-bp target fragment. baranagar west bengalWebJun 15, 2015 · Construction of recombinant pcDNA3-HBsAg-p30-ROP Fresh recombinant plasmid pcDNA3-p30-ROP2 was extracted. Restriction enzyme digestion was performed … baranakhsWebMar 8, 2024 · Plasmid construction. pcDNA3.1 (+) vector was purchased from Thermo Fisher Scientific. STXBP5-AS1 DNA was obtained by PCR using cDNA, Pfu DNA polymerase and synthetic oligonucleotide primers incorporating restriction sites. PCR products were ligated into the pcDNA3.1 (+) vector according to the manufacturer’s … baranailt road