We narrowed to 695 results for: prot

...Green Fluorescent Protein Resources on Addgene.org Browse our Fluorescent Protein Collection Find Plasmid...sensors. Given the expanding palette of fluorescent proteins and the availability of powerful imaging technologies...technologies, the combination of multiple fluorescent protein sequences to develop diverse fluorescent reporters... reporters can be used to detect structure and protein localization, as well as cellular processes like...modules used in this method include fluorescent proteins, promoters, enhancers, polyadenylation signals... created, each containing a unique fluorescent protein with minimal spectral overlap between them (Table...In each construct, 3 copies of the fluorescent protein were fused in-frame to a tethering partner, which...

...neuroscience-related proteins, but we recently added several broadly useful antibodies for detecting protein tags.  ... saturating mutagenesis on 21 positions in the protein sequence of the predecessor GEVIs (called ASAP ...Kalium channelrhodopsins (KCRs), from a fungus-like protist Hyphochytrium catenoides (Govorunova et al. 2022...  And when you think of protein tags, what are the first tags that come to mind? I’m willing to guess ...EQKLISEEDL, that can be incorporated into your protein of interest and recognized by reliable antibodies...derived from the C-terminal region of the human proto-oncogene c-Myc. Since the generation of the first...Myc tag has become one of the most commonly used protein tags - a quick search for ‘Myc tag’ in the Addgene...

...such as Rep or other initiator proteins, they do not require host proteins for replication and can maintain...as recognition sites for replication initiation proteins. These iterons vary in their sequence and number...oris interact with various RNA polymerases and proteins from bacterial hosts. In a plasmid with multiple... (ori1, ori2, ori3) and replication initiation proteins (RepA, RepB) necessary for replication in various...has its own replication initiation factors and proteins independent of the host. Host range depends on...can replicate in different bacteria and express proteins within different host organisms. pBBR1-based plasmids...Pasteurella multocida, Porphyromonas gingivalis, Proteus mirabilis, Providencia spp., Pseudomonas spp., ...

...paper or protocol. If you have questions while reading these materials (maybe a step in the protocol you don...ask for a copy of the protocol if you prefer to take notes on a printed protocol, or maybe you would like...resources to help you get started!) Are there protocols you can familiarize yourself with? If your bench...bench mentor hasn’t sent you papers or protocols yet, don’t be scared to ask for some! You can email or...or “It would be great if you could send me the protocol we’ll be using next week so I can familiarize ...bare minimum should be taking notes on general protocol steps and other tips and tricks your mentor shares...next few days or weeks), look over your notes and protocol(s). Is there enough information there for you ...

...fluorescent proteins is only possible at low copy number ranges, resulting in low fluorescent protein level....system to express one of the three fluorescent protein (XFP) genes in a transgene. When multiple copies...2018) (Figure 1). Once introduced, fluorescent proteins were best imaged after tissue clearing with SeeDB2.... For more details on this procedure, find the protocol here.   Tips and troubleshooting your Tetbow...expression levels. Clearing large tissues - Fluorescent proteins are very stable in SeeDB2. However, SeeDB2 is ...High-resolution fluorescence imaging using SeeDB2." Bio-Protocol 8 (2018): e3046. Livet, Jean, et al. "Transgenic...strategies for combinatorial expression of fluorescent proteins in the nervous system." Nature 450.7166 (2007)...

...tagging of every protein in the C. elegans genome. Although they tested fluorescent protein fusions, this...even easier - now, one can generate a fluorescent protein fusion, transcriptional reporter, and loss-of-function... allele in just one injection step! The entire protocol takes about 2-3 weeks but requires less than eight...SEC). To make it possible fluorescently label proteins in C. elegans, they inserted the SEC into a synthetic...synthetic intron between a fluorescent protein (FP) and a 3xFLAG tag (see construct schematic below.) ...this synthetic intron, and only the fluorescent protein and 3xFLAG-tag sequences are inserted into the ...plasmid to create fluorescent and 3xFLAG-tag fusion proteins. The repair template and Cas9-sgRNA plasmid is...

...allows you to ask questions like: “Where does my protein of interest live within a cell,” “Does this disease...where they bind to their targets, most commonly proteins. You can then use fluorescent microscopy to look... multiplexing is that you can ask how multiple proteins relate to one another in the same sample. For ...other antibodies. But, once you do find the right protocol, multiplex IF can give you impactful and beautiful...manufacturer’s recommendations.  As you optimize your protocol, always remember to include controls to show that...experiment will have a significant impact on the protocol you use and the ultimate outcome of your assay...share some of their results and corresponding protocols. Other researchers may also have tested an antibody...

...where a fluorescent protein is inserted into one of the loop regions of any G-protein-coupled receptor (...Fluorescent Proteins 101: Fluorescent Biosensors Resources on Addgene.org Fluorescent protein plasmids ...circularly-permuted GFP into an intracellular loop of various G-protein-coupled receptors. Image adapted from Ino et al...calcium reporter Near-infrared (NIR) fluorescent proteins promise better performance for in vivo use and...indicator using small monomeric NIR fluorescent proteins (miRFP670nano and miRFP720) and the Ca2+-sensing...structure-guided engineering of potassium binding proteins, they developed a family of fluorescent potassium...Introducing mutations and using potassium-binding protein homologues from various bacteria led to sensors...

...repair proteins fused to CGBEs The David Liu lab also experimented with adding different proteins to the... binding motif protein X-linked (RBMX) (Koblan et al., 2021). Several of these proteins had additive effects... (2) a complementary DNA sequence, and (3) a protospacer-adjacent motif (PAM) on the opposite strand. ... cytosine transversion editors. New DNA repair proteins fused to CGBEs Instead of using UNG, the Wei Leong... CBE with other BER components: the DNA repair protein rXRCC1, or the DNA binding and lyase domain of ...Mycobacterium smegmatis, termed UdgX, they found three proteins whose fusion improved the CGBE: DNA polymerase...improving C → G editing. Instead of fusing DNA repair proteins to the base editor, they added the pioneer factor...

...Vesicular stomatitis virus G glycoprotein; Broad tropism envelope protein; Pseudotyped to alter infectivity...(viral Gag-Pol): Gag is a structural precursor protein, and Pol is a polymerase. Envelope gene (may be...packaging (Gag, Pol, and Env), the isoforms of these proteins, as well as the viral LTRs, differ. As a result...plasmids, such as VSV-G (Vesicular Stomatitis Virus glycoprotein), however, may be used across both systems.  ...pseudotyping.   Figure 1: Genome of a prototypical gamma-retrovirus.    Frequently asked questions...infectivity. Packaging Gag Precursor structural protein of the retroviral particle containing Matrix, Capsid...and Nucleocapsid components. Pol Precursor protein containing Reverse Transcriptase and Integrase ...

...psychrophilic proteins. The primary problem we wanted to overcome was inefficient protein folding at low...psychrophilic proteins. As the figure below shows, E. coli expressing the chaperones protein resulted in...Following up its 2018 iGEM debut - centered around a prototypical methane biosensor - IISER Bhopal is back in ...enzymes for the host The expression of recombinant proteins for industrial or research purposes requires stable...example E. coli, carrying a plasmid expressing a protein of interest). Mesophilic microbes are highly preferable... standard conditions. However, one of the many proteins which are not compatible with regular mesophilic... suboptimal conditions. This means recombinant proteins can aggregate or precipitate out as insoluble ...

...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...

...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...

...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 ...

...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...