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We narrowed to 130 results for: yeast expression

Showing: 61 - 80 of 130 results

  1. Don’t FRET: Bimolecular Fluorescence Complementation Makes Visualizing Protein-Protein Interactions Easy

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    Blog Post
    ...Right Connection. Cellular expression Whether BiFC constructs are expressed transiently by transfection... independent of PPIs. Overexpression can be assessed by comparing the expression levels between BiFC constructs...open-reading frame (ORF) libraries have been generated for yeast (Kim et al., 2019) and fruit flies (Bischof et al... steps: fusion protein construction, cellular expression, and quantification (Kerppola, 2006, 2013). These...this can also be perturbed by supraphysiological expression levels. BiFC quantification This is where BiFC...cannot be used as negative controls because their expression level, stability, and subcellular localization...Kerppola, 2006). Here, one of the endogenous PIPs is expressed at increasing levels alongside your BiFC constructs...

  2. A Look Back at One Year of Plasmid Sharing for COVID-19 Research

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    Blog Post
    ...synthesized SARS-CoV-2 expression plasmids available for many expression systems: bacterial, yeast, and mammalian...C-terminal deletion in the spike protein to improve expression and pseudotyping efficiency. This plasmid was...enzymes. Plasmids in this toolkit include ones for expressing MMLV reverse transcriptase and Taq polymerase...-CoV infection and deposited plasmids for the expression of cathepsin S, L, and B. This became a great...human cells. Addgene’s mammalian target plasmids express these and other targets such as TMPRSS2, FURIN,...and become upregulated during viral infection.  Expression of SARS-CoV-2 viral proteins In the early weeks...Krogan lab at UCSF generated lentiviral vectors expressing SARS-CoV-2 open reading frames and identified...

  3. New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression

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    ... that, when expressed, enable cells to be visualized under ultrasound imaging. Expression of ARGs results...in bacteria, expressed poorly at body temperature (37oC) and were too burdensome to express in situ. As...vesicle expression in E. coli is easy: simply grow cells in Lennox LB (10 g/L tryptone, 5 g/L yeast extract... gene expression in deep tissue. However, not unlike the first-generation bARGs, poor expression and acoustic...Increasing expression and decreasing genetic footprint We took a cue from the native expression patterns...copies of gvpA needed to increase expression. We were able to express Anabaena GVs in mammalian cells ...microscopy just one day after induction of expression… but longer expression leads to greater amount of gas vesicles...

  4. Plasmids 101: Knockout/Knock-In Plasmids

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    ...constructed plasmids can be used to replace genes in yeast, mice, or Drosophila through homologous recombination...disrupted (knocked out) but the inserted GFP is expressed (knocked in). As seen in the above example, you...control of an endogenous promoter, you can use expression GFP to track cells participating in development... in intronic regions, this gene will still be expressed. You'd first screen for this specific recombination...Off Resources on Addgene.org Find Mammalian Expression Vectors Find Cre-Lox Plasmids Learn More about...

  5. Plasmids 101: Choosing an Antibiotic Resistance Gene

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    Blog Post
    ...bacteria that express the Tn5 element, but thankfully, most competent cells do not express this factor....of the plasmid, isolate individual plasmids, or express a gene contained in a plasmid. During the propagation...confound the goal of your experiment — to grow, express, or isolate plasmids. The presence of an antibiotic...The Sh ble gene provides zeocin resistance by expressing a protein that directly binds to zeocin, preventing...antibiotic — it is usable in eukaryotic cell culture, yeast, bacteria, and even plants! The zeocin resistance...never entirely detoxified, even when Sh ble is expressed (Trastoy et al). Thus, it may not be a good choice...2012). SHuffle, a novel Escherichia coli protein expression strain capable of correctly folding disulfide...

  6. GCE4All: Making Genetic Code Expansion Accessible

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    Blog Post
    ...techniques being developed. In the mid-aughts, the first yeast GCE paper was published, followed soon by GCE done...that you need to worry about getting high enough expression of your protein of interest. If that wasn’t enough...protocol is available now. It is for recombinant expression of phosphoserine-including proteins in E. coli...

  7. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

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    Blog Post
    ...specific) control of DNA recombination and gene expression can be advanced by the coordinated use of fellow...means to spatiotemporally control FLP and Cre expression have been developed. Read on to learn more about...ability to invert or ‘flip DNA’, is derived from the yeast S. cerevisiae (Gronostajski and Sadowski 1985). ...SV40 Large T nuclear localization sequence) and expressing it from a CAGGs promoter. However FLPe is still...the DNA sequence of interest can lead to poor expression of genes within that sequence and should therefore... in neuronal networks Double inverted vectors expressing light inducible ion channels (plasmid # 20298... specific manipulations in vivo, FLEx vectors expressing the light activatable ion channel under the control...

  8. Multiple Plasmids at a Low Price: Kits from Addgene

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    Blog Post
    ...of promoters to facilitate gene expression in bacteria, yeast, mammalian cells, insects, and even Xenopus...Process with pCri Cell-free protein expression kit Cell-free expression systems have been improved for both...comparison of different cell-free expression systems; the Cell-free expression test kit represents a set of...benchmarking of cell-free expression systems, and tested for product integrity, expression level, and aggregation...BIOFAB Plasmid Set pCri System Cell Free Protein Expression Kit FREQ Seq PUF Assembly Kit Below you’ll find...it easier to produce the constructs you need. Expression vectors within this kit contain a variety of ...Xenopus oocytes. The expression vectors additionally contain various combinations of protein tags (His, Avi...

  9. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

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    Blog Post
    ...their small size, nanobodies can be synthetically expressed off of plasmids in bacteria, and they can target...comparison to other display systems using phage and yeast (109-1010 members). This selection step involves...screens are subcloned (using FX cloning) into the expression vector pSb-Init, which attaches a Myc-tag and...sybodies can be subcloned into pBXNPH3 or pBXNPHM3 expression plasmids for the production of tag-free proteins...

  10. Getting the Most from Your Lentiviral Transduction

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    Blog Post
    ...often employed to establish cell lines that stably express a gene of interest. While the general approach ...quiescent cells do not take up nucleic acid or express transgenes as well as actively dividing cells resulting... unlike infections with larger microbes such as yeast, fungi, or bacteria, mycoplasma can be extremely...compete with host cells for nutrients and can alter expression of receptors, ion channels, and growth factors...on a variety of factors such as the transgene expressed, the viral storage buffers used, and other experimental...will be able to overcome differences in receptor expression. It's all in the mix Even before the lentiviral...infection significantly alters microarray gene expression profiles.” Biotechniques 35.4 (2003). 812-4. ...

  11. Antibodies 101: Single Chain Fragment Variables (scFvs)

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    Blog Post
    ...because they can be expressed in bacteria, while mAbs generally require mammalian expression systems. scFvs...scFvs in mammalian, yeast, plant, and insect cells, they are most often expressed from a plasmid in bacteria...Ahmed et al., 2012). One challenge of bacterial expression is the proper formation of the disulfide bond...requires an oxidizing environment to form. When expressed in the non-oxidizing cytoplasm of bacteria, scFvs...more of these approaches are often used for scFv expression in bacteria:  targeting to the oxidizing periplasm...to allow for proper disulfide bond formation.  expression in strains of bacteria that are redox mutants...mutants so they have a more oxidizing cytoplasm. expression in the presence of molecular chaperones that help...

  12. CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?

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    ...for expression in different species and cell types including (but not limited to) bacteria, yeast, plants...modify the genome but still want to affect gene expression? dCas9 can be used as a platform to deliver various...activation domains to the same genetic locus via co-expression of an epitope tagged dCas9 and antibody-activator...Validated gRNAs for Your Next CRISPR Experiment Cas9 expression systems Addgene carries a variety of Cas9 containing...systems can be found here “CRISPR 101: Mammalian Expression Systems and Delivery Methods”. Validating your...

  13. Plasmids 101: Screens vs. Selections

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    Blog Post
    ...interest (or the whole genome), they knock down gene expression temporarily. RNAi libraries are especially useful...assayable with CRISPR. Another method, popularized in yeast, are temperature sensitive screens. These involve...

  14. 10 Basic tips for mammalian cell culture

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    ...microscope will help you identify bacterial and yeast contaminants. Set up a routine to split cells Don...LinkedIn.   References 1. Yin, R., et al. "Gene expression profiling analysis of bisphenol A-induced perturbation...cytoskeletal disorganization, apoptosis, and gene expression disregulation on HeLa cells." Journal of applied...

  15. Plasmids 101: Modular Cloning

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    ... multiple parts transcribed together. They can express one or more gene(s), and typically begin with a...strategies are not available to those working with yeast or plants. Using MoClo, or better yet, a pre-made...

  16. Technique: Probe Phage Genomes for Host Binding Proteins

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    ...homology-based searches return no candidates. Our strategy: Express random fragments of the phage genome and probe ...plate) across 20 fresh selective plates. Step 2: Express random phage DNA fragments in E. coli Now that ...and exposed to IPTG in order to induce protein expression. Cells on the membranes will then be exposed ...sleeping) point for this experiment. Find Bacterial Expression Vectors in the FX Cloning System Kit  Step 3:... and growing them up overnight. Assuming they expressed well, the proteins encoded by the plasmids in ...the membrane itself or to any “sticky” proteins expressed by your library. After blocking for 1 h at room...at room temperature. This will allow any RBPs expressed on the membrane to bind their target cells, pulling...

  17. Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection

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    ...genes close to native expression levels. However, it is advisable to first express the fusion protein from...strategy (Sheridan, 2002). Avoiding protein over-expression artifacts The introduction of DNA that encodes...of protein within a cell and may lead to over-expression artifacts. This is an important downside of the...characterisation of fluorescent proteins in budding yeast." Scientific reports 9.1 (2019): 2234. PubMed PMID...

  18. Microbiology Resources

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    Collection
    ...Vectors - Barbier Lab Cloning and Expression Bacteria and yeast expression vectors : FX cloning System - ...oleaginous yeast Y. lipolytica - Rossignol Lab Yeast Gateway kit : 285 vectors for expression, markers,...and biosafety regulations. Looking for E. coli or yeast plasmids? Addgene has a wide variety of tools for...Sets of plasmids for fluorescence, cloning and expression, and synthetic biology. CRISPR Resources : Find...plasmids intended for use together (e.g. cloning and expression systems) and therefore supplied as one item. ...Fluorescent Proteins FRET-based biosensors : E. coli expression - Pertz Lab Bacterial Fluorescence Labeling :...- Geertsma and Dutzler Labs EasyClone 2.0 Yeast Toolkit - Borodina Lab Golden GATEway cloning system -...

  19. Transgenic Organisms, Cas9 Gene Drives, and Appropriate Safeguards

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    ...spread exclusively through populations that already express Cas9 from an unlinked locus, while a Cas9+sgRNA...efficiently and reversibly bias inheritance in wild yeast. bioRxiv 013896 (2015). doi:10.1101/013896. bioRxiv...methods are easy to implement and have been tested in yeast5. Ecological confinement involves performing experiments...

  20. Fluorescent Protein Guide: Empty Backbones

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    Collection
    ...PA-GFP-KanR - Yeast Expression pFA6a-PA-GFP-kanMX6 - Yeast Expression pFA6a-link-yEPAGFP-CaUra3 - Yeast Expression... N1 - Mammalian Expression mVenus C1 - Mammalian Expression pKT0090 - Yeast Expression mVenus-pBAD - Bacterial...pFA6a-link-tdTomato-SpHis5 - Yeast Expression tdTomato-N1 - Mammalian Expression tdTomato-C1 - Mammalian Expression tdTomato-pBAD...Mammalian Expression pFA6a-link-yomRuby2-SpHis5 - Yeast Expression mRuby2-N1 - Mammalian Expression mRuby2...Mammalian Expression pTagRFP657-C1 - Mammalian Expression pFA6a-link-yoTagRFP657-Kan - Yeast Expression smURFP...pFA6a-link-yomKeima-CaURA3 - Yeast Expression mKeima-Red-N1 - Mammalian Expression mKeima-Red-C1 - Mammalian Expression dKeima...pBad-EBFP2 - Bacterial Expression EBFP2-N1 - Mammalian Expression EBFP2-C1 - Mammalian Expression EBFP2-pBAD - ...
Showing: 61 - 80 of 130 results