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We narrowed to 130 results for: yeast expression

Showing: 1 - 20 of 130 results

  1. K. phaffii: Rising to the Occasion

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    ...model yeast strain and discuss why you may want to consider a yeast off the beaten path.   Yeast as a ...Resources on Addgene.org  Yeast-mediated cloning CRISPR plasmids: Yeast Yeast Synthetic Biology Plasmids...what about the other yeasts – in the biomedical world, are there even other yeasts? In this blog we will... frequently used yeast strains: S. pombe (fission) and S. cerevisiae (budding). Yeast make for an excellent...useful traits. The “biotech yeast” You’re probably wondering if the “biotech yeast” is S. pombe or S. cerevisiae...non-pathogenic yeast that operates as an obligate aerobe. It’s also a methylotropic yeast, meaning it can...and biotech industries as a protein expression workhorse. This yeast can also give more protein bang for...

  2. Plasmids 101: Yeast Vectors

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    ...previously described. Why do scientists use yeast vectors? Yeast are eukaryotes and thus contain complex ...facilitate simple gene replacement/mutation. Yeast expression plasmids used in the lab typically contain...between E. coli and yeast cells. To be useful in the lab, the vectors must contain a yeast-specific origin...selection in yeast cells, in addition to the bacterial ORI and antibiotic selection markers.  The yeast origin...within a yeast vector determine how the plasmid is replicated and maintained within the yeast cell. These...episome. The four main types of yeast plasmids are defined below: Yeast Integrating plasmids (YIp): These...the yeast chromosome. As the name suggests, these vectors can replicate independently of the yeast chromosome...

  3. Tips for Screening with Yeast Two Hybrid Systems

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    ...Browse Our Yeast Expression Vectors Find Bacterial Two Hybrid Plasmids Read our Blog Post About Yeast Vectors...years, improvements to yeast transformation protocols also helped establish the yeast two-hybrid (Y2H) system...Figure 2a) (4). These fusions are then co-expressed in a yeast strain that also contains a reporter gene...doesn’t function in yeast! Many false negatives stem from or are exacerbated by expression of target proteins...gene fusions for analyzing gene expression in Escherichia coli and yeast. Methods Enzymol. 1983;100:293...activation. Vary the expression levels of bait and prey proteins - overexpression can lead to false interactions...During this period, proteins were successfully expressed as fusions that retain their individual activities...

  4. Plasmids 101: Protein Expression

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    ...specifically interleukins. Yeast expression systems Yeast are a great expression system to generate large...plant, yeast and cell free expression systems. Overall the general strategy for protein expression consists...which expression system to use depends on several factors: The protein you are trying to express How ...system. Mammalian expression systems Mammalian cells are an ideal system for the expression of mammalian proteins...for mammalian expression utilize viral promoters (SV40, CMV, and RSV) for robust expression post-transfection...insect expression systems; baculovirus infected and non-lytic insect cells. Baculovirus expression systems... level recombinant protein expression. These systems enable high expression of very complex, glycosylated...

  5. Five Popular Model Organisms

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    ...spatiotemporal expression of a gene of interest. This type of control over gene expression is possible in...optimally control gene expression in specific cells of the fly (Pfeiffer et al., 2010). Yeast (Saccharomyces .... Like human cells, yeast DNA is packaged into chromosomes and about 23% of yeast genes have a counterpart...homolog involved in yeast cell division (Pray, 2008). Scientific discoveries in yeast can then can be further...lewy bodies” in the brain. Yeast models of PD, like those that overexpress the toxic protein α-synuclein...Saccharomyces cerevisiae) Yeast, one of the simplest eukaryotic organism, is a commonly used model organism in scientific...same kind we use in breads and other baked goods! Yeast is cheap, simple and easy to work with as they can...

  6. Pathways Over Time Plasmids Engage Students in Functional Genomics Research

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    ...Figure 1) from the two yeast species into the pYES2.1/V5-His yeast overexpression vector. This plasmid ...at Addgene Resources on Addgene.org Find Yeast Expression Vectors Check Out Our Molecular Biology Reference...gene in the reference species. We have been using yeast models (3, 4) to answer questions about the evolution...involved in methionine synthesis (5). The budding yeast, Saccharomyces. cerevisiae, makes a great reference...students test whether or not proteins from the fission yeast, Schizosaccharomyces pombe, have the same function...as their counterparts in S. cerevisiae. The two yeasts are thought to have diverged from a common ancestor... or S. cerevisiae. Students then determine if expression of the plasmid-encoded MET gene allows the transformed...

  7. Tag Your Favorite Yeast Genes with Ease

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    ...proteins (FPs) to a yeast gene of interest. These fluorescently tagged genes are expressed under native conditions...that are codon optimized specifically for expression in yeast. These tagging vectors are based on previously...homology—in frame, of course—and start tagging! Yeast-optimized fluorophores for imaging Lee S, ...authors recommend optimal FP combinations for use in yeast imaging, categorized by specific filter sets and...experimental output requirements. Select from these yeast-optimized fluorophore tagging vectors for your single...has built a complete collection of yeast tagging modules with each possible combination of protease site...collection of plasmids for genome modification in yeast developed by Dr. Pringle’s former lab at UNC Chapel...

  8. Plasmids 101: Shuttle Vectors

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    ...neither, depending on the yeast species and the goal of the experiment. Yeast shuttle vectors have these... selection is especially common in yeast experiments because yeast tend to spontaneously develop resistance...vectors in fast-growing bacteria first. In fact, most yeast vectors and many mammalian vectors are designed ...used across species: replication, selection, and expression. Replication It’s usually not enough to get your...genomes. Eukaryotes are a bit more complicated. Yeast vectors may contain an autonomously replicating ...these replication sequences for their yeast targets as well as a bacterial ori for prokaryotic growth. Since...this approach requires compatible cell lines that express viral machinery. Alternatively, some slow-growing...

  9. Addgene-Seeding Labs Plasmid Grant: Accelerating Science Globally - 5 Scientists, 4 Countries & 3 Continents!

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    ...using yeast integration vectors (see for example this yeast cloning kit) she intends to make yeast capable...developing yeast as source for dietary supplements! Development of a bacterial expression system for ...the light‐inducible bacterial expression system pDawn to clone and express newly identified proteins and...project the expression of isolated bacterial proteins can be achieved using a bacterial expression system ...different CRISPR applications in plants.     Transgenic yeast as source of dietary supplements Dr. Mrinalini ...focuses on molecular biotechnology for improving expression of nutritionally and therapeutically significant.... In this project she aims to create transgenic yeast as an alternative source of essential omega-3 fatty...

  10. Plasmids 101: The Promoter Region – Let's Go!

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    ...Description Expression Additional considerations CMV General expression mRNA Strong mammalian expression promoter...neuronal/CNS expression. Modulated by calcium and calmodulin. GAL1, 10 General expression mRNA Yeast adjacent...mammalian EF1a promoter. GDS General expression mRNA  Strong yeast expression promoter from glyceraldehyde 3...lower expression than U6. May have better expression in neuronal cells. U6 small RNA expression shRNA...some cell types. EF1a General expression mRNA Strong mammalian expression from human elongation factor ...give consistent expression regardless of cell type or physiology. SV40 General expression mRNA Mammalian...tet transactivator is used. UAS General expression mRNA Yeast promoter containing Gal4 binding sites, ...

  11. Plasmids 101: Modular Cloning Applications and Kits

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    ...looking to express high levels of their gene of interest in yeast should also take a look at the Yeast GoldenBraid...used in bacterial and cell-free expression systems, mammalian cells, yeast, flies, and even plants! In this.... coli and/or cell-free systems. Protein expression in yeast MoClo is also for eukaryotes! The technology...optimize protein secretion and expression in the biotech workhorse yeast — K. phaffii (formerly known as... difficult in alternative expression systems such as bacteria. If your yeast experimental goals are more...conditional gene expression, multiplexed CRISPR, and chromosomal integration of larger expression cassettes ...chromosomal integration and gene expression, the system includes a toolkit to express recombinant proteins within...

  12. Hot Plasmids and Viral Preps - May 2021

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    ...aminoacyl tRNA synthetases Nanocompartments for yeast bioproduction pathways The CRISPR corner New ready-to-use... (B) Mapping antibody-escape mutations with the yeast display library. From Starr et al Science., 2021...that the SARS-CoV-2 spike RBD is displayed on the yeast cell’s surface. The Bloom lab used the library to...protein cage provides simple enzyme protection in yeast bioproduction pathways By Gabrielle Clouse Claudia...novel nanocompartment for transporting proteins in yeast by using murine polyomavirus virus-like particles...this method, the major capsid protein VP1 is co-expressed with a fusion protein containing VP2C and a protein... freely into or out of the protein compartment. Yeast with this encapsulated enzyme produced 20% more ...

  13. Choosing Your Perfect Empty Backbone

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    ...binding or folding assays. Yeast vectors There are several types of yeast expression vectors. Consider your...as close as possible to the basal expression level, induce expression at an opportune time or go all out...gene by transforming wild-type yeast with an integrating vector expressing a selection cassette with 5′ ...purposes Addgene has a large collection of yeast expression vectors with various markers and promoters... following the expression pattern of a defined promoter region, ectopically expressing YGOI in a pattern...for overexpressing YGOI in an isolated, controlled environment. In most cases bacterial expression vectors... in molecular biology to isolate, multiply, or express the insert they carry in the target cell. These...

  14. Hot Plasmids April 2018 - Protein Degradation, Nanoscopy, FIRE-Cas9, and Yeast Expression Tools

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    ...of gene expression in yeast Article contributed by Tyler J. Ford Listen to the Synthetic Yeast Transcription...encoding synthetic yeast transcription factors that can independently control expression of your genes of...independent control of gene expression, Aranda-Díaz et al transformed yeast with both a GAL4/estradiol ...variation, or noise, in reporter expression. To do so, one places reporter expression under the control of two...factor 1 controls expression of transcription factor 2, which directly controls expression of a reporter ...contain changes in expression of TRIM21-associated genes, indicating that TRIM21 overexpression does not aberrantly...physiological gene expression regimes that cannot be easily achieved with classical gene overexpression and knockout...

  15. Rosella: A Fluorescent pH-Biosensor for Studying Autophagy

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    ... to study autophagy in yeast Rosella is targeted to the cytosol when expressed without a signal sequence...analyzing autophagy of cytosol and organelles in yeast cells. Autophagy (Greek for “self-eating”) is induced...it is slow. Alkaline phosphatase activity: In yeast, the PHO8 gene encodes a vacuolar alkaline phosphatase...monitor autophagy, the Pho8Δ60 mutant protein is expressed. Pho8Δ60 localizes to the cytosol in an inactive...a group of genes that affect autophagy (ATG) in yeast. Atg8p (“p” stands for protein) associates with ...structures, autophagosomes and autophagic bodies in yeast. A GFP-tagged version of LC3, the mammalian Atg8p...autophagy by tracking what’s being transported to the yeast vacuole (i.e. cytosol, mitochondria), rather than...

  16. 27 Hot Plasmids from 2016

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    ...for in-vivo expression and plasmids optimized for dCas9-VPR expression in drosophila and yeast. Chavez ... HEK293FT cells expressing a single gRNA targeting a single gene. The gRNA expressing cells were then ... to test multiple expression systems before they achieve the appropriate expression level for their gene...create protein expression constructs fused with epitopes or fluorophores, generate expression vectors with... lab’s own pAc-sgRNA-Cas9 insect expression backbone, which expresses the gRNA from a Drosophila U6:2 ...genetic optimization of industrial yeast strains. However, industrial yeast strains are more difficult to ...model organisms. As long as it was expressed in a cell that also expressed auxin-sensitive TIR1, any protein...

  17. Plasmids 101: Repressible Promoters

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    ...to be expressed. These permit exquisite control of gene expression and tracing of gene expression across... basal expression. Co-expression of repressor QS along with QF leads to intermediate expression from QUAS...UAS system isolated from yeast. In this system, UAS basal promoter expression is low but is activated ...transgene permits directed expression of that gene in cells that also express GAL4. Usage of this system...UAS to examine the expression of reporter genes, or to combine reporter expression from one promoter with...basal state, and co-expression of the inducer QF can increase QUAS driven expression by ~3,300-fold in ... Shen. “Controlling gene expression with the Q repressible binary expression system in Caenorhabditis ...

  18. Hot Plasmids - August 2020

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    ...easier to clone, express, and purify compared to antibodies. Nanobodies are easily expressed in the cytosol...#140702) The pMAK461-64 plasmids were expressed in SHuffle T7 Express competent E.coli (NEB) using IPTG induction...approaches rely on continual expression of both dCas9 and the gRNA. This expression can vary significantly ...up here.  Here's what you'll find in this post: Yeast-based protein evolution Fluorescent tools for mammalian...from the viral service Faster catalysis after yeast-based protein evolution By Joanne Kamens Mateo Sanchez...to optimize another. The authors have created a yeast-based directed evolution approach with some notable...involves selection in the cytosol (instead of on the yeast extracellular surface) which enables tracking of...

  19. Pushing the Limits of DNA Assembly

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    ...Producing yeast expression cassettes Traditionally, generating expression cassettes for yeast transformation...point mutations Cloning sgRNAs Production of Yeast Expression Cassettes Generating constructs with multiple...linearized cassette, ready for transformation into yeast that same day.  The field of synthetic biology has...

  20. What's New in CRISPR - March 2020

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    ...demethylation Optogenetic repressors of gene expression in yeast Megan McClean’s lab generated an optogenetic...CRISPRa depended on continued expression of dCas9 and gRNAs whose expression varies depending on transgene...research! This time: Optogenetic repression in yeast Base editing CRISPRi in Caulobacter crescentus Multiplexed...optogenetic toolkit that controls repression of gene expression in Saccharomyces cerevisiae. The system consists...the nucleus where dCas9-Mxi1 can repress gene expression. In the dark, the NLS is inaccessible and the...of CRISPRi in Caulobacter cells for ctrA gene expression using various dCas9. Image adapted from Guzzo...lab developed a CRISPRi system to downregulate expression of specific Caulobacter genes using a catalytically...
Showing: 1 - 20 of 130 results