...their genomes. Unlike plasmids used for expression in yeast and bacteria, plasmids used for stable transfections...Your Perfect Plasmid Backbone Read More: Plasmids 101: Antibiotic Resistance Genes Plasmids 101: Origin...Although plasmids do not naturally exist in mammals, scientists can still reap the benefits of plasmid-based...straightforward and the resultant cells can either express the plasmid DNA transiently (similar to bacteria) or ...reporter is unique to the cell, is expressed from the plasmid, and can be assayed conveniently. A direct..., it is sufficient for the transfected plasmid to be expressed transiently. Since the DNA introduced in...components expressed in trans within the cell for effective replication. Cell lines expressing the Epstein–Barr...

...Should I Use? Plasmids 101: Green Fluorescent Protein (GFP) Fluorescent Protein Guide Expression Vectors from...localization, or protease protection. Thus far Plasmids 101 has covered GFP and its related fluorescent proteins...box of tags and fusion proteins to aid in the expression and purification of recombinant proteins. Tags...stability activity, can be readily purified using plasmids found in this paper (available at Addgene).   ...   plasmids 101...Biochemists and molecular biologists who need to overexpress and purify proteins can face any number of technical... some of the hurdles to overcome in order to overexpress a recombinant protein? It is not generally in...

.... pBLADEONLY: This plasmid contains only BLADE and is based on the pTRC99a plasmid. It does not include...promoter. If you already have a pBAD33-based plasmid expressing your gene of interest, all you have to do...used a strain transformed with pBLADE expressing superfolder GFP instead of mCherry under the PBAD promoter...-inducible AraC dimers in E. coli) that turns expression on and off with blue light.   Inspired by @UriAlonWeizmann...AraC and other promoter systems, head over to our Plasmids 101: Inducible Promoters article.) Modifying AraC...wild-type AraC, it still reached the level of expression needed for most applications.   Figure...reporter downstream of the PBAD promoter. To use this plasmid with your gene of interest, you would replace mCherry...

... into the brain of Cre-expressing mice to trigger recombination and expression of a transgene (Lin et .... GAL4 expressed in the fly binds to the UAS binding motif, driving downstream gene expression. attP and...effector is expressed. In cells where the stop cassette remains, the effector is not expressed and the cells...components and expression is controlled by the upstream recombination outcome. Find SPARC plasmids at Addgene...controlling gene expression in flies Resources on Addgene.org Find fluorescent protein plasmids at Addgene...the system or using chemical inducers of gene expression (del Valle Rodríguez et al., 2011).  Genetics... construct. A predictable number of cells will express the effector or retain the stop cassette following...

...phosphatidic acid probe GFP-PASS. B) Time series of HEK 293T cells expressing plasma membrane-targeted... of katanin to that anchor. So, you’ll need to express both the EB3N-VVDfast-iLID anchor module and the...module, which are also available as lentiviral plasmids if you want to make a stable cell line. This design...specific area at the plasma membrane (Figure 2). Just express the plasma membrane-localized iLID along with your...Opto-RhoGEF design and activity. B) OptoTIAM (magenta) expressed in a monolayer of endothelial cells, showing cell...either CRY2 or CIB1 to your protein of interest and express the other separately, then use blue light to generate...developing embryo is accessible for injection of plasmids and other materials.    Figure 3: Optogenetic...

...Find the plasmids from this publication at Addgene. Multichromatic control of gene expression in Escherichia...these two systems can be coexpressed for sophisticated control of gene expression. An E. coli light-sensitive... by Tabor’s lab, making light-controlled gene expression in E. coli easier and more robust. Synthetic...engineer new pathways with sophisticated gene expression controls. Most cell types do not respond to low-intensity...via phosphorylation, promoting transcription of a GFP reporter. Subsequent exposure to red light rapidly...light-responsive system is spread across three plasmids, with the response regulator encoded in the genome...et al. condensed the system to two streamlined plasmids. The utility of these TCSs was limited by a few...

... blot for the myc-tagged protein GFP-smFP-myc expressed from Plasmid 98926. See the Applications section...of the new plasmids, antibodies, and viral preps in the repository through our hot plasmids articles. ...Think we missed something hot? You can pitch a Hot Plasmids section (antibodies and viral preps welcome!) ... you can also find several other constructs in plasmid form. Fig. 1: Summary of the approach and...JEDI-2p viral preps here (and here!)Find JEDI-2p plasmids here!  Liu, Zhuohe et al. Sustained deep-tissue...B) Cortical slice of HcKCR1-EYFP and tdTomato expressed layer 2/3 neurons in mouse. C) Action potential...voltage traces of action potentials in a HcKCR1-expressing mouse neuron without (top) and with (bottom) ...

... (ARG) plasmids to better monitor gene expression in vivo using ultrasound. These ARG plasmids contain...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles ...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need... recent plasmid deposit please sign up here.      Listen to this episode of the Hot Plasmids podcast ...pXPR_053. They expressed this construct in bone marrow stem cells from Cas9-expressing mice, and used ...responses in C. elegans. Find the split BoTN/B plasmids at Addgene. Liu Q et al., Neuron. 2019. PubMed... microbe travels through a host organism, the expression of some genes can change depending on the location...

... FRET tools, double dimerization GFPs, and circular permutation GFPs. Hopefully we will have new tools...Plant Kit makes it really easy to clone genes for expression in plant cells. Sylvester and I chose the 96 ...that are not used so much but have comparable expression levels to things people do use a lot. The problem...susceptible to a lot of pathogens. It’s also easy to express genes in, which makes it a good laboratory workhorse...simultaneous complementation, with a knockout and expression construct on the same piece of DNA. We do a lot...Quick, Versatile Plant Transgenesis with GreenGate Plasmids  ...

...without mCherry and GFP fluorescent controls. Alternatively, you can order individual plasmids from the library...Addgene.org Plasmid Pooled Libraries Pooled library amplification Zhang Lab CRISPR Plasmids The Human TFome...factors (TFs) act as the gate keepers for gene expression, turning transcription on and off by binding ...binding proximal to their target genes. Since gene expression patterns determine everything from stem cell differentiation...(aka the MORF Collection) and an accompanying expression atlas, including over 3,500 human TF isoforms...TF library, with each cell receiving only 1 TF expression construct. Cells were cultured following a differentiation...from Joung et al.)    This atlas linked TF expression to differentiation outcomes as well as global...

..., 13xMyc, GST, or GFP). Longtine et al.(3) describe a complimentary set of plasmids for use in S. cerevisiae... artifacts sometimes associated with plasmid-based overexpression. Tim Formosa, at the University of Utah...Addgene distributes several ready-to-use, modular plasmids, combining fluorescent tags, epitope tags, protease...interest. These fluorescently tagged genes are expressed under native conditions and allow scientists to...FPs that are codon optimized specifically for expression in yeast. These tagging vectors are based on ...). Additionally, Dr. Formosa has deposited six plasmids with a multiple cloning site in place of the epitope...Jürg Bähler have deposited a large collection of plasmids for genome modification in yeast developed by ...

... is now complete! pAAV-CAG-GFP (plasmid 37825) and pAAV-hSyn-EGFP (plasmid 50465) are now available as...directs strong, ubiquitous expression. This is a Cre-dependent switch that expresses nuclear mCherry in the...entire AAV inventory. Our new AAVs include: EGFP-expressing AAV for serotype testing Calcium sensors and...testing AAV, which are small (20 ul) samples of EGFP-expressing AAV packaged in various serotypes. This collection...pAAV-hSyn-DIO-EGFP (50457-AAV5): The Synapsin promoter directs broad, neuronal expression. AAV pCAG-FLEX-EGFP-WPRE...pCAG-FLEX-EGFP-WPRE (51502-AAV5): The CAG promoter directs ubiquitous expression. pAAV-mDlx-NLS-mRuby2 (99130...directs expression in forebrain GABAergic interneurons. pOTTC1032 - pAAV EF1a Nuc-flox(mCherry)-EGFP (112677...

...specimens, such as whole mouse brains expressing green fluorescent protein (GFP) (6).   The Dodt and Frank Bradke...cranial nerve afferents labelled with a AAV8 expressing eGFP under the human ubiquitin C promoter. Anatomical...subpopulation of neurons are brightly labeled with GFP (e.g. Thy1-YFP-H mice).  However, we recently developed...compared to transgenic animals (17).  These viral plasmids are available through Addgene.  The combination...University of Miami Miller School of Medicine. Find plasmids from the Lee Lab here. Pantelis Tsoulfas is an...University of Miami Miller School of Medicine. Find plasmids from the Tsoulfas lab here.     References: H...

... size of 12–15 kDa Ease of cloning and expression from plasmids, due to only needing one open reading ...highlighted back in 2015, where sdAbs were used to create GFP scaffolds for transcriptional activation.  sdAbs ... because sdAbs are so amenable to recombinant expression (and because maintaining a llama facility is ...interesting work done developing transgenic mice that express camelid variable domains (aka nanomice), which ...domains of heavy chain only antibodies) (B) Nanomice express hybrid heavy-chain only antibodies that are the...Membrane Proteins More resources from addgene.org  Plasmids encoding nanobodies at Addgene Sybody Generation...

...accelerate your research.   Why distribute virus? Plasmid distribution is booming, but, over the years, many...list of what’s available or suggest additional plasmids with the Viral Service Suggestion Form. Lentivirus... to knockout a gene, replace it, modulate its expression, or perform a genome-wide screen, we’ve got viruses...also have lentivirus for delivering control shRNAs, GFP, and rtTA from the Tet-on system. The first two can...experiments, while rtTA can be used to activate expression of a gene under the control of the tet operator...page and includes western blots as well as other expression data. If you have any questions about the Viral...Uses and Overview AAV: A Versatile Tool for Gene Expression in Mammals Additional Resources on Addgene.org...

... single protein is expressed, with the intent of purification, it will be expressed along with all other...other cellular proteins within the expression system (bacteria, yeast, mammalian, etc.). To individually...about that protein that no other protein in the expression system has. This is where affinity tags come ...peptides which do not naturally exist in most expression systems that can be added to the N or C terminus...on: The protein you are trying to purify The expression system If you want the tag to remain on your ...MBP tag, although large, can actually increase expression and solubility of the tagged protein (Fox and... (FLAG, Myc, etc.) and fluorescent protein tags (GFP, mCherry, etc.). Fluorescent tags are primarily used...

...Resources on Addgene.org Find Biosensor Plasmids Find Optogenetics Plasmids Visit the Fluorescent Protein Guide...associated publication for the specifics of your chosen plasmid. Types of biosensors When choosing a biosensor...biosensors as an example. Many calcium biosensors include GFP fused to calmodulin (CaM) and the chicken myosin ...translocation. Targeting your biosensor Before expressing a biosensor in eukaryotic cells, think about ...about the best place for the biosensor to be expressed. Are you looking at an intracellular or extracellular...

...Description Expression Additional considerations CMV General expression mRNA Strong mammalian expression promoter...lower expression than U6. May have better expression in neuronal cells. U6 small RNA expression shRNA...promoter, which is necessary for getting the plasmid express our gene of interest. This element is responsible...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...  Commonly used in expression systems for Drosophila. Polyhedrin General expression mRNA  Strong insect...Constitutive  Commonly used in expression systems for insect cells. CaMKIIa Gene expression for optogenetics mRNA...

...selection. Expression is driven by a fairly strong EF1alpha promoter. We have verified expression of correctly-sized...2 virus-human interaction network in which we expressed 26 of the 29 SARS-CoV-2 viral proteins in human...the viral protease, Nsp5. Our set also includes a GFP construct as a control. For easier portability, all...Western Blot for 25 constructs, and confirmed expression via mass spec for two of the remaining viral ...going on there!).  Find the Krogan Lab SARS-CoV-2 plasmids (Note: The Nsp6 construct is undergoing QC and... has been inundated with requests to share our plasmids with other researchers. We have been excited to...Gordon) have been quite busy shipping and sending plasmids to all who request them -- in locations all over...

...high-copy plasmid-borne chromoproteins lost color overnight in liquid culture, though expressing the chromoproteins...for allelic exchange Figure 3: pTOX plasmids expressing amilCP and tsPurple genes. Image: Lazarus...coli BioBrick plasmids (meffRed, eforRed, asPink, spisPink, scOrange, fwYellow, amilGFP, amajLime, cjBlue...contrast, the widely used green fluorescent protein (GFP) gives a bright green fluorescence when exposed to...at the cells on the plate. For example, if your plasmid contains a gene encoding a blue chromoprotein, ...colonies to determine which cells have taken up the plasmid. Of course, other follow up methods like sequencing...gfasPurple) (Figure 2). Figure 2: Chromoprotein expression from bacteria on agar plate. Image: Liljeruhm...