We narrowed to 883 results for: prot

...Proximity Ligation Assay for Detecting Protein-Protein Interactions and Protein Modifications in Cells and Tissues...the antibody sequence or that rely on strong protein-protein interactions with other peptides, like an Fc-binding...non-canonical amino acids or peptide tags. 5) Strong protein-protein interactions with other peptides. Created with...quantitative immuno-PCR. Nat Protoc 2:1918–1930. https://doi.org/10.1038/nprot.2007.267 Stoeckius M, Hafemeister...Tissues in Situ. Curr Protoc Cell Biol 89:e115. https://doi.org/10.1002/cpcb.115  Mueller BM, WrasidloO...

... ccd operon, codes for the antitoxin protein (CcdA) that protects the cell against the toxic CcdB. Cells... plasmid CcdB killer protein: ccdB gene mutants coding for non-cytotoxic proteins which retain their regulatory...Technique Find Plasmid Cloning Protocols Check Out All of Our Protocols Additional Web Resources OpenWetWare's...during cell division. ccdB codes for the toxic protein (CcdB) that acts as a DNA gyrase poison, locking...Addgene Blog for a variety of useful resources and protocols! We invite feedback from all members of the research...7926841. 4. Bernard, P., et al. “The F plasmid CcdB protein induces efficient ATP-dependent DNA cleavage by...Couturier, M. “Cell killing by the F plasmid CcdB protein involves poisoning of DNA-topoisomerase II complexes...

... targeted genetic edits. Cas9 is a DNA binding protein that can be directed to virtually any genetic locus...that is complementary to the target site, or protospacer, in the genome. Native Cas9 has two catalytic...which cleaves one strand of DNA upon binding the protospacer. The resulting double strand break (DSB) stimulates...catalytic domains can be mutated to inactivate the protein’s nucleolytic capability, and this nuclease-null...transcriptional activators or repressors. These fusion proteins retain the ability to recognize and bind to DNA...proposed the concurrent use of “orthogonal” Cas9 proteins derived from different bacterial species, each...distinct gRNA that allows users to decide which Cas9 protein is directed to which target site (1). Although ...

...to modified proteins called histones. We can detect modified histones with “GPS for proteins,” ChIP-seq...the genome. Aneesh's research interests include proteomics, machine learning, and visualization for big ... compute coordinates for genetic elements like protein, RNA, and DNA (Table 1). As with GPS in the real...techniques Technique What It Locates ChIP-seq Proteins (for our example later, histones) ChiRP-seq ...For details on DNA compaction and the structural proteins that make it possible, see the appendix DNA is... you run a ChIP-seq experiment (think “GPS for proteins”) for NANOG, an essential transcription factor...epigenomics: H3 – one of NANOG’s associated histone proteins K4 and K27 – locations of the amino acid lysine...

...is formed by a complex of two proteins: the large (L) catalytic protein and its cofactor VP35. VP30 is...viral proteins (compared to viral infection). Therefore, the roles/functions of individual proteins can ...transcribed in the cell when the appropriate viral proteins are provided in trans (see Other Important Components... they do not encode all of the necessary viral proteins. Minigenome systems have been developed for many...genome and transcribes viral genes. Four viral proteins are required for viral genome replication and ...in 5). The RNA genome is encapsidated by the nucleoprotein, NP. NP associates with the viral polymerase...approach. First, this would prevent the study of the proteins independently from each other (see Uses of Minigenome...

...pieces: a single protein, computationally described as a fusion of four Cas7 proteins and a putative Cas11...significant!) protein engineering. They combined Prime editing with an integrase via protein fusions, using... into E. coli to express and then test for any proteins that could cut RNA or DNA via a CRISPR mechanism...black pellet's DNA yielded a new CRISPR effector protein they named Cas7-11. Testing showed that Cas7-11... III-B systems. Though it was composed of many protein subunits, it was nonetheless a new subtype. Cas7...-11 had combined all the domains into a single protein, shedding the complexity of its ancestors. They... Cas11-like protein, and a guide RNA. The enzyme has dual activities: processing pre-CRISPR RNA into mature...

...users to search for protocols in protocols.io, and then link to a selected protocol and/or import and convert...powerful example is the integration with protocols.io, the protocol sharing repository. As illustrated in... possible to re-export a protocol which has been edited back to protocols.io. Thus, RSpace becomes part...management software, and method repositories like protocols.io. These tools are essential to the workflows ...enhances the sharing and reproducibility ecosystem protocols.io makes possible. Connectivity to open source ...scientific sharing Resources on Addgene.org Find protocols of molecular biology techniques Find plasmids ...

...both strands. Similar to how Cas proteins require a PAM (protospacer adjacent motif) near the target DNA...Transposon-Associated TNPB system. In Springer protocols handbooks/Springer protocols (pp. 245–260). https://doi.org/...genome editing toolbox. However, both of these proteins are quite large — comprising approximately 1000... thought to be evolutionary ancestors of Cas12 proteins. Functionally, while Cas9 contains two nuclease...changes based on the species of origin of the TnpB protein, much like how PAM sites change for different Cas...(Molla et al., 2021), which require fusing Cas proteins with additional enzymes or domains (increasing...accessible genome engineering. More elaborate protocols are available in our earlier publications (Karmakar...

...identification of plant compounds and insecticidal proteins to the development of renewable biotechnology ...system pDawn to clone and express newly identified proteins and enzymes. These compounds will later get tested... nutritionally and therapeutically significant proteins. In this project she aims to create transgenic...desaturating fatty acids. Identification of Insecticidal Proteins Linh Bao Ton (M. Sc.), Nong Lam University, Vietnam...and analysing the insecticidal potential of Cry proteins in order to manage insect pests and to support...this project the expression of isolated bacterial proteins can be achieved using a bacterial expression system...vector and the pOKD plasmids. The recombinant Cry proteins will then be screened for toxicity against local...

...your protein in the cell or purify it – and you do not have a specific antibody against the protein available... you can always add an epitope tag or a fusion protein. Mammalian vectors One of our most popular mammalian...The addition of an in-frame fluorescent fusion protein will allow you to visualize YGOI in vivo (try pcDNA3...6xHis or GST) to affinity purify the expressed protein with a specific resin (e.g. 6xHis containing pDEST-HisMBP...pDEST-HisMBP). After purification the protein can be released from the resin and used for various binding... collection of vectors carrying several fusion proteins. Pick your promoter, selection cassette and visualization...pattern that allows the function of the encoded protein to be assayed (e.g. pPD95_86 and pPD57_56), or ...

...Viruses are simple: a genome packaged in a protein shell (Taylor, 2014). They’re so simple that we can...own machinery to replicate the viral genome and proteins. This is, for the intrepid researcher looking ...structure While all viruses have a genome inside a protein shell known as a capsid, some also have a lipid...can either follow central dogma (gDNA to RNA to proteins) or require an intermediary DNA replication step...transcribed into mRNA, directly translated into proteins, or, in the case of retroviruses, reverse transcribe...transcribed into mRNA before being translated into proteins. RNA viruses replicate their genomes using a variety...ability. The remaining particle, which retains the protein coat (and envelope), has a “gutted” genome with...

...and using RNA scaffolds to assemble proteins in vivo. Nature protocols. 7(10) 1797-1807 (2012). PubMed. ...CRISPRi) model consists of a catalytically dead Cas9 protein (dCas9) that lacks endonuclease activity coupled...into a secondary structure that can bind specific proteins via aptamer domains. Combined with dCas9, this...enzymes for violacein, a distinctive pigmented protein. By altering the transcriptional programs, the ...scRNAs all individually utilize the same dCas9 protein. By using an inducible promoter to modulate dCas9...cell fate decisions. By using an orthogonal dCas9 protein that will still recognize guide RNA but with a ...effectively change metabolic flux. The dCas9 protein and orthogonal variants can be used as regulatory...

...DNA Quantification Protocol Recovering Plasmid DNA from Bacterial Culture Protocol ... plasmid DNA from bacterial cultures is a core protocol in molecular biology. And while kit-free plasmid...quality/low quantity) – Bacterial transformation protocols require such small amounts of DNA (often on the...process and include things like genomic DNA, RNA, proteins, salts, ethanol, or endotoxins. The most common... kits — which to choose? Plasmid purification protocols can be broken into a few broad stages: lyse the...environmental impact. Using non-silica-based purification protocols requires less plastic and avoids the use of more...have suggested modifications to their normal kit protocols.  Yeast plasmids – If you’re extracting plasmids...

...fluorescence intensity of a tagged protein. SunTag has two components: 1) a protein of interest that’s tagged ... are often the building blocks for engineering proteins. So let’s get acquainted with scFvs. What is an...antibody.  The ability to combine scFvs with other protein domains, much like lego pieces, makes them better...antibodies to be used as building blocks for engineered proteins. This flexibility is largely due to scFvs being...antibodies. scFvs are often genetically fused to other proteins, which has led to their application for basic ... HA-binding scFv that’s fused to a fluorescent protein such as GFP. The HA frankenbody works just like... Whitlow M (1988) Single-chain antigen-binding proteins. Science 242:423–426 . https://doi.org/10.1126...

...separate enzymes that are proteolytically processed from the larger protein: PR (protease), RT (reverse transcriptase...their life cycle to continue. They rely on a viral protein called reverse transcriptase to convert their RNA...typically encodes the vesicular stomatitis virus G protein (VSV-G), which facilitates viral entry into the... integrated into the host genome via the viral proteins packaged during production (read our blog post...into a large complex of both viral and cellular proteins and trafficked to the nucleus. Since the integrase...transgenes/shRNAs If you want your cells to express a protein or nucleic acid for only a limited time, IDLVs ...Lentiviral Guide and our lentiviral production protocol.  One thing to keep in mind is that titering these...

... purification protocol involving iodixanol gradients. The current purification protocols for isolating...accumulate inside the producer cells and purification protocols often describe a process that begins with cell... also contain other cellular molecules such as proteins or nucleic acids. This conjunction of vectors ...Since the development of the exo-AAV isolation protocol by Maguire and colleagues, other researchers have...cells also contain cellular macromolecules (e.g. proteins, nucleic acids) which possibly improve transduction...association of AAV particles with membrane vesicles protects them from antibody recognition and inactivation... contexts, such as the delivery of therapeutic proteins to the brain which requires crossing the blood-brain-barrier...

...of a protein, a yes-no plot is the best-case result. For example, you have introduced a GFP protein into...adjusted voltage, cells that did not receive the GFP protein are most likely to be clustered at signals below...not! When you are looking to study the level of protein expression, instead of its presence or absence,...These smears tend to occur when you are looking at proteins that are expressed regularly in cells and might...The orange peis the baseline expression of the protein of interest. Created with biorender.com. The... the orange peak is a baseline expression of a protein while the green (top) has an upregulated expression...Priyamvada Prathima has a research background in protein mutagenesis and cancer immunotherapies. She is ...

... Quadros et al. recommend ribonucleoprotein (RNP) complexes of Cas9 protein and the separated guide RNA...using long ssDNA donors.” Nat Protoc. 13 (2018) (1):195-215. DOI: 10.1038/nprot.2017.153 PubMed PMID: 29266098... example, large insertions, like a fluorescent protein knock-in, suffer from low efficiency when a standard...desired insertion feature (e.g., a fluorescent protein), you can amplify the desired region using primers...the commonly used cassettes, such as fluorescent proteins (EGFP, mCherry etc.), recombinases (Cre, Flp etc...You can also use Addgene plasmids to purify Cas9 protein. The table below contains some of our favorites...alleles using long ssDNA donors and CRISPR ribonucleoproteins.” Genome Biol. 18 (2017) (1):92. DOI: 10.1186...

...editors that can be reconstituted in vivo to form a protein identical in sequence to the unmodified enzyme....Genetically encoded voltage indicators (GEVIs) are proteins that detect and output the voltage changes in ...inhibitory rhodopsin (Ace2N) with the red fluorescent protein (mScarlet) with either a seven or eight amino acid...generated knock-out and knock-in mutants, determined protein expression levels from different promoters, and...half with PhyB or PIF6, phytochrome-like fusion proteins from the plant Arabidopsis thaliana. In the presence... light or PCB is no longer present, the fusion proteins separate, inactivating Cre. This is critical for...truncated Phytochrome B (PhyBΔ) and the active protein binding domain of Phytochrome B interacting factor...

...be transformed into Alphaproteobacteira, Betaproteobacteria, and Gammaproteobacteria species isolated from...self-assembling protein (SAP) such as a bacteriophage shell protein or a bacterial microcompartment protein. The ...fluorescent proteins into one particle within the cell, the lab fused the red fluorescent protein (RFP) to... (DCDC), relies on the assembly of fluorescent proteins into one bright particle that segregates into ...