Ef1a promoter vs cmv promoter

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This promoter configuration takes advantage of the natural bidirectional activity of the PGK promoter, and this divergent configuration is both stable and resistant to silencing in embryonic stem (ES) cells, trophectoderm stem (TS) cells, and extra embryonic endoderm (XEN) stem cells 1. A promoter is a region of DNA where transcription of a gene is initiated. Promoters are a vital component of expression vectors because they control the binding of RNA polymerase to DNA. RNA polymerase transcribes DNA to mRNA which is ultimately translated into a functional protein. In thalli transformed with vectors containing either the SV40 or CMV promoter, lacZ presence was confirmed by histological staining 2 and 3 days postbombardment. In thalli transformed with the vector containing the CaMV 35S promoter, lacZ presence was confirmed by histological staining 1, 2, 3 and 5 days post-bombardment. May 10, 2019 · Trying to spread knowledge and make it easily accessible to everyone, everywhere. The WT CMV promoter is known to be poorly expressed in some stem cell lineages, although the factors responsible are unknown . This phenomenon may be specific for the CMV minimal promoter, as the CMV enhancer is functional in hematopoietic stem cells in concert with other promoters . These data suggest that a specific repressive factor may act ... • Dual promoter format for high level marker expression • Can be used as either plasmid or packaged lentivirus • CMV and MSCV promoter options • D-Luciferin reagents also available for molecular imaging in vivo Nov 16, 2015 · The FOS promoter caused significantly higher transcriptional activity in glioma cell lines than all alternative promoters with the exception of CMV. The FOS promoter showed 13.9%, 32.4%, and 70.8% of the transcriptional activity of CMV in three glioma cell lines (U87, U251, and U373). In my experience, the CMV promoter is stronger than the EF1a promoter in 293T cells, but I never quantified this, so I'm only making an 'educated' guess. Is it the same cDNA which is cloned in the vectors you are using? The pLV-cDNA vector is a self-inactivated (SIN) vector carrying a deletion in the U3 region of the 3’ LTR, which eliminates the promoter activity of LTR. The self-inactivating deletion provides an additional level of safety by preventing the integrated viral genome from generating full-length lentiviral RNA. Sep 28, 2020 · Other promoters (e.g CMV) may work well in cell culture but can sometimes get silenced in vivo. When expressing FPs, you should always try to use a strong ubiquitous promoter (e.g. EF1A or CAG) if you can. If you have to use a tissue-specific promoter, you should also choose a strong one when possible. Secreted Gaussia Luciferase Lentivirus CMV Promoter or EF1a Promoter The Secreted Gaussia Luciferase Lentivirus are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to be transduced into almost all types of mammali See full list on blog.addgene.org strong in all the cell types, with EF1A being slightly stronger in some cell types and slightly weaker in others. SV40 promoter is also fairly strong, though generally somewhat weaker than EF1A and CAGG. While there is cell type to cell type variability for all the promoters, CMV promoter is the most variable, being very strong in some cell Plasmid pLV-EF1a-IRES-Puro from Dr. Tobias Meyer's lab is published in Nat Cell Biol. 2016 Dec;18(12):1311-1323. doi: 10.1038/ncb3438. Epub 2016 Nov 14. This plasmid is available through Addgene. Could anyone tell me the difference between CMV and CMV-IE promoter? Top. Log in or register to post comments; Thu, 06/26/2008 - 11:42 ... Since the promoter region drives transcription of a target gene, it therefore determines the timing of gene expression and largely defines the amount of recombinant protein that will be produced. Many common promoters. like CMV, EF1A, and SV40 promoters, are always active and thus referred to as constitutive promoters . In addition to one-promoter vectors, which carry the multiple cloning site downstream of the CMV promoter, we offer the option of a second expression cassette downstream of the MCS to express the puromycin or neomycin resistance gene or copGFP as a reporter under the control of a constitutive promoter. introduced Ptet-1 [1] is based on a minimal promoter fragment derived from the CMV immediate early pro-moter. Various alternatives were introduced over the years, like those further truncating the CMV minimal promoter of Ptet-1 [12-15], constructs based on the * Correspondence: [email protected] 1EUFETS GmbH, 55743 Idar-Oberstein, Germany posttransfection (Fig. 1A). The CMV promoter showed the highest activity, while K19 0.7-kb promoter had the least activity, in A5 cells. The human EF1a promoter exhibited the highest activity of all mammalian pro-moters, only about threefold lower than that seen with the CMV promoter. The two K18 promoters tested I'll add my voice to the thread to offer up Wootric (http://www.wootric.com) as another online tool to consider for a Net Promoter system implementation. We're ... The integrated provirus has two LTRs, and the 5' LTR normally acts as an RNA pol II promoter. The transcript begins, by definition, at the beginning of R, is capped, and proceeds through U5 and the rest of the provirus, usually terminating by the addition of a poly A tract just after the R sequence in the 3' LTR. Aug 20, 2020 · 2 nd position: promoter: BamH1/Nde1 2nd gene CDS: Nde1/Cla1. NB: Nde1 enzyme requires 7 base pairs in after the restriction sequence for efficient cutting. Also beware CMV promoter has an internal Nde1 site so this site will not be unique when working with CMV containing constructs. The first one has a CMV promoter, the second one a U1 promoter. ... As an example, the pEF vector available from Sigma, as well as most EF1 vectors on addgene have a 1172 bp EF1a promoter sequence. 156 commercial promoter jobs available. See salaries, compare reviews, easily apply, and get hired. New commercial promoter careers are added daily on SimplyHired.com. The low-stress way to find your next commercial promoter job opportunity is on SimplyHired. There are over 156 commercial promoter careers waiting for you to apply! A promoter is a region of DNA where transcription of a gene is initiated. Promoters are a vital component of expression vectors because they control the binding of RNA polymerase to DNA. RNA polymerase transcribes DNA to mRNA which is ultimately translated into a functional protein. A suitable replacement for the pCBh promoter is a CAG promoter. The CAG promoter consists of three genetic components that will help drive expression of proteins within a hESC. Its components include a Cytomegalovirus (CMV) enhancer, a Chicken-β-actin promoter, and a Rabbit β-Globulin Splice acceptor gene. Previous studies have shown that the ... Jun 09, 2018 · Transcriptional promoter: Promoter is a – • Region of DNA that control the transcription of a particular gene. • It locate upstream of the genes. • Recognizing site for RNA polymerase (sigma subunit). Good promoter should be: • Strong promoter: Accumulation of expressed protein up 10 to 30% or more of the total cellular protein. posttransfection (Fig. 1A). The CMV promoter showed the highest activity, while K19 0.7-kb promoter had the least activity, in A5 cells. The human EF1a promoter exhibited the highest activity of all mammalian pro-moters, only about threefold lower than that seen with the CMV promoter. The two K18 promoters tested Apr 11, 2019 · We have recently published a paper about CMV enhancer/promoter in which we discussed in detail the CMV enhancer/promoter and further improved the transcriptional activity of the CMV enhancer by changing the natural NF-κB binding sites in this enhancer into artificially selected NF-κB binding sequences with high binding affinity (Wang et al ... into the pcDNA3.1-cmv-FVIIIDB, the pBS-HCR-1-2 vector was digested with the enzymes EcoRV and XhoI to release the HCR-1-2 fragment. This fragment was treated with klenow enzyme to form blind ends and cloned into the vector pcDNA-cmv-FVIIIDB. This vector was previously digested with MluI (restriction site located upstream to the CMV promoter) The promoter of this ’housekeeping’ gene exhibits a strong activity, higher than viral promoters such as SV40 and RSV promoters [1] and, on the contrary to the CMV promoter, yields persistent expression of the transgene in vivo [2]. InvivoGen has cloned the EF-1α promoter region of different species, among them the chimpanzee and rat. Apr 20, 2005 · EF1a – a „strong“ native mammalian promoter active in most or all tissues. Not as strong as viral promoters like CMV. Tends not to get silenced (like CMV does). Sometimes the EF1a core promoter with CMV enhancer is used. Mammalian Promoter Vector Set plasmid vectors for molecular cloning; Synonym: cloning vector, expression vector, molecular cloning vector, plasmid, plasmid vector, vector; find Sigma-Aldrich-PP2356 MSDS, related peer-reviewed papers, technical documents, similar products & more at Sigma-Aldrich. SV40 promoter is also fairly strong, though generally somewhat weaker than EF1A and CAGG. While there is cell type to cell type variability for all the promoters, CMV promoter is the most variable, being very strong in some cell types (e.g., 293T and CMMT) and rather weak in others ( e.g.,MRC5. Actived: 5 months ago The pLV-cDNA vector is a self-inactivated (SIN) vector carrying a deletion in the U3 region of the 3’ LTR, which eliminates the promoter activity of LTR. The self-inactivating deletion provides an additional level of safety by preventing the integrated viral genome from generating full-length lentiviral RNA. Could anyone tell me the difference between CMV and CMV-IE promoter? Top. Log in or register to post comments; Thu, 06/26/2008 - 11:42 ... Promoter: CX3CR1 (chemokine receptor CX3CR1) Transgene: Cre-ERT2 Notes: BAC transgenic using rat CX3CR1 promoter to express Cre recombinase in microglia. LE-Rosa26Tm1(LSL-Cas9)Ottc. Request: RRRC #833 Promoter: CAG (synthetic hybrid construct consisting of the cytomegalovirus (CMV) enhancer fused to the chicken beta-actin promoter) Mar 13, 2019 · Mean mCherry fluorescence (driven by the activation-inducible promoter) was normalized to the copGFP signal (driven by the constitutive EF1a promoter), in order to factor in the differences in fluorescence associated with altered cell size and general increase in transcription background following cell activation. The pRL-CMV vector contains the cDNA encoding Renilla luciferase (Rluc) and the CMV enhancer and early promoter. After transfection, cells were incubated at 25°C for 6 h, transferred to 1.5-ml microcentrifuge tubes and placed in thermostatted waterbaths at either 36.5°C (heat shock) or 25°C (control) for 5 min, transferred to a 25°C ... The integrated provirus has two LTRs, and the 5' LTR normally acts as an RNA pol II promoter. The transcript begins, by definition, at the beginning of R, is capped, and proceeds through U5 and the rest of the provirus, usually terminating by the addition of a poly A tract just after the R sequence in the 3' LTR.