...should be properly disposed of following your institution’s hazardous waste procedures. Last Update: February...February 7, 2022 Protocol Video Watch this instructional video to learn how to affinity purify recombinant...for your columns, refer to the manufacturer’s instructions. Attach the Gravitrap column to a clamp on a...

... also use a nucleotide-based molecular insert (inosine) instead of a regular nucleotide for broader pairing...

...of plasmids in your purified DNA. This protocol explains how to isolate a single bacterial colony by streaking...plasmids or bacteria need to be grown at 30 °C instead of 37 °C. This is often true for large unstable...

...Envelope plasmid, and Transfer plasmid encoding your insert of interest Reagent Preparation DMEM Complete (... optimized for each transfer plasmid. Pro-Tip Endotoxins can inhibit transfection; therefore, plasmid ...

...ensure that you are in compliance with your institution’s biosafety regulations. Last Update: February... °C. See below for other Reagent Preparation instructions. Wipe down all pipettes and equipment with 10%...

...purification and isolation of cells, organelles, lipoproteins, and macromolecules. Importantly, iodixanol ...of contaminants in each fraction. * AAV capsid proteins VP1, VP2, VP3; M protein marker. Image adapted...

...cloning cylinders. Workflow Timeline This protocol begins with a stable cell pool. For details, see our protocol...homogenized cell solution to 990 µL complete medium. Instead of transferring 0.125 µL, now transfer 100 times...

...you have a NanoDrop, follow the manufacturer's instructions for the spectrophotometer specific to your lab...

...Extraction Kit . Always follow the manufacturer's instructions. Note: It is usually important to determine ...

...ensure that you are in compliance with your institution’s biosafety regulations. Workflow Timeline Day...

...µg of DNA into 500 µL of Opti-Pro SFM. Pro-Tip Endotoxins can inhibit transfection; therefore, plasmid ...

...the liquid and avoid touching the pipette to the inside of the tube. Only the tip should be in the liquid...

...into polypeptide chains that make up the primary structure of proteins. Most proteins are then modified...systems. Insect expression systems Insect cells can also be used to produce complex eukaryotic proteins with...Researches can visualize proteins in vivo by tagging them with fluorescent proteins to study localization...localization or purify proteins to study their structure, interactions and functions. Proteins can also be purified... to manipulate DNA), or in medicine (e.g. insulin). Proteins, unlike DNA which can be relatively easily...are two types of insect expression systems; baculovirus infected and non-lytic insect cells. Baculovirus...genes integrated into the insect cell genome. Both of these types of insect expression systems can be ...

...       Insert-specific primers: Insert-specific primers are designed to anneal to an insert-specific... your insert or if the insert is in your plasmid Have to make a new primer pair for each insert Backbone-Specific...the insert, insert orientation, and the sequences of the junctions between the plasmid and insert DNA....negative clone without the insert. This type of primer pair can tell you if the insert is the correct size and...orientation of the insert. Orientation-specific primers: If you need information about insert orientation,...of the insert. One member of this type of primer pair anneals to a sequence flanking the insert and one...anneals to the insert. A simple way to create this type of primer pair is to mix-and-match insert-specific ...

...reporters as well as secretory proteins or proteins targeted to membrane sub-domains. It has also been used to... reporter mRNA, which contains the tethering sites for the fluorescent proteins and an inducible promoter...and the RFP proteins in the nucleus. During the first round of translation, the GFP proteins that are bound...the nucleus (thanks to a NLS on these proteins). The RFP proteins, however, remain bound to the mRNA since...the Wyss Institute have extended the transactivating capabilities of CRISPR/dCas9-fusion proteins and have...engineered hook proteins fused to a streptavidin core that are able to retain reporter proteins fused to the...possible protein domains containing these properties, they generated monoclonal antibodies against them, fused...

... open a plasmid (backbone) and insert a linear fragment of DNA (insert) that has been cut by compatible... allow your insert to be placed into the backbone in the proper orientation. For instance, if you were...enzyme if your insert is flanked on both sides by that enzyme’s restriction sites, but the insert can then ...used. Follow the manufacturer’s instructions.   2. Isolate Your Insert and Vector by Gel Purification ...separate your backbone away from any inserts cut out of it and your new insert from any overhangs cut off of ...these stains see the Bitesize Bio Blog and their associated manufacturers websites. These stains require...Your Insert into Your Vector In the ligation step, you mix your purified, cut backbone and insert in a...

...fused to two proteins, here proteins Y and Z, that are known to interact. When  proteins Y and Z interact...interacting proteins; nearby proteins are biotinylated when and where the two bait proteins interact, but...interact or from proteins that are just in the proximity of the bait proteins. Instead of pinpointing specific...restored, and nearby proteins are labeled with biotin. For labeling, BirA* fusion proteins are expressed in...activity is restored, and nearby proteins are labeled with biotin. When proteins Y and Z part ways, BirA* biotinylation...the biotin labeling remains even if the interaction with the BirA* fused bait proteins ceases. Lastly, Split-BioID...provides a candidate list of proteins that are part of the bait proteins’ interactome. Further characterization...

.... The Moffat laboratory maintains a website for the TKO library that contains up-to-date information and...(3) but its portability to other bacterial strains remains seriously limited, as prior optimizations are...ligase to biotinylate proximate proteins in the cell. These candidate proteins can be enriched via biotin ...of fusion proteins, requires less biotin, and shows enhanced labeling of proximate proteins. Overall BioID2...optimization of industrial yeast strains. However, industrial yeast strains are more difficult to genetically...which “bait” proteins are tagged with Protein A-Renilla luciferase and the “prey” proteins are tagged with...of RNA-binding proteins identified by editing Identifying targets of RNA-binding proteins (RBPs) has traditionally...

... next insert. Repeat as needed for more inserts. When you are finished entering your gene/insert data,...cloning a gene/insert into it. Empty backbones often contain epitope tags or fusion proteins that are appended...within your insert (or targeted by a gRNA/shRNA) and select the species that the gene or insert sequence ...include the total size of the insert and the GenBank ID associated with your insert. Pro Tip! If the correct...of the insert (when cloned into the vector backbone). If your plasmid has more than one insert, click .... For instance, if your plasmid is designed to allow you to produce large amounts of your insert in E....it. If your plasmid has multiple inserts, enter details for each insert individually.   Figure 9: Add ...

... either side of the expected insert. If the plasmid contains your insert, you will see two bands: one ...another enzyme that cuts within the insert. If the plasmid contains the insert, you will see two bands but if...the plasmid contains the insert and no PCR product if the plasmid does not contain the insert. If you’re...but it won’t tell you whether the vector contains the gRNA insert or not. A blue-white screen, however, ...sized bands depending on whether the plasmid contains your insert of interest or not. For example, choose ...the other representing the insert. If the plasmid doesn’t contain the insert, you will just see the vector...PCR to detect your insert, either by using primers that are specific to your insert or by using primers...

...isolating proteins from a specific compartment. If your proteins are nuclear- or DNA-binding proteins, you ...step: protein degradation, incomplete lysing/insoluble proteins, and protein aggregation. Protein degradation...gel box. If overheating remains an issue, which can happen with large proteins, try lowering the voltage...transferring are smaller proteins going completely through the membrane or larger proteins not transferring completely...sizes for different proteins: try using a 0.2 µm (psi) membrane for small proteins (< 15 kDa) and a 0.45...which works better for small and medium proteins. If your proteins are compatible with electroblots/dry ...migration of smaller proteins through the membrane while increasing the ability of proteins to stick to the...

...primary antibodies conjugated to fluorescent proteins, instead of the more common chemiluminescent methods...or β-mercaptoethanol.  Denaturing proteins To denature your proteins, you'll want to use SDS and either...-life; can alter proteins due to high pH Bis-tris 6–400 kDa MES for proteins <40 kDa 180 V, ...weight proteins Long running time   It’s important to note that Bis and Tris gels run proteins in different...They work well for proteins of most sizes and are worth considering if your proteins of interest vary wildly...protein you are expecting.  Fluorescent proteins Fluorescent proteins, while more expensive, allow for multiplexing.... Protein preparation Before you can blot your proteins, you’ll need to lyse and denature them. If you...

... the Institute for Protein Innovation (IPI), a Boston-based, nonprofit protein research institute, partnered... closed states, integrins are inactive, with low affinity for ligands. When integrins interact with extracellular...the role of specific integrins in cellular processes.   Figure 1: Integrins can shape into three...recognition site for one subfamily of integrins, the RGD-binding integrins. Six of the 11 IPI antibodies contain...Targeting integrins in the lab The Ha and Springer labs recently showed RGD-binding integrins demanded ...RGD-binding integrins, individually and in combination, showing that among the three αV integrins expressed...Integrins are essential cell surface receptors involved in complex signaling pathways and the linkage...

...wages, overtime tracking? What are institutions doing? Institutions are now preparing to comply with the...resource on what institutions are actually doing (using the list of 340 institutions with postdocs in .... Most institutions suggest that postdocs should be paid according to the National Institutes of Health...In 2014, 89% of institutions had a minimum salary policy: of those, 51% of institutions set their postdoctoral...should be on the lookout for what their institutions, or institutions they're thinking about applying to, ...salaries will have to double at some institutions; and that institutions which can more easily provide bridge...scientists and those in technical roles. Therefore, institutions are responsible for ensuring that either all...

...integral membrane proteins and membrane associated proteins (i.e. GPI-anchored proteins), which are more... National Institutes of Health and he is currently an Associate Professor at Albert Einstein College of...tagging to detect proteins expressed from cloned genes: deletion mapping functional domains of Drosophila ...Fluorescent Proteins Browse Our Fluorescent Protein Pages Find the mOX Fluorescent Proteins at Addgene  ...to many microscopy experiments with fluorescent proteins (FPs) localized to compartments other than the... a resident endoplasmic reticulum (ER) protein instead localizes to the cytoplasm? Or more troubling, ...approximately 40% of human (and most eukaryotic) proteins localize to chemically distinct subcellular environments...

...were at institutions focused only on tracking hours, and 22% of postdocs were at institutions where no... of some public institutions) to cancel plans to raise salaries. What are institutions doing after the...planning unionization at their institutions. One clear message that institutions have sent, with the lack of...their institution is doing from Future of Research over social media, not from their institution), is ...raise were: postdocs at institutions like Stanford and the Whitehead Institute, where salaries had already...street from these institutions) are raising salaries. Postdocs at the Salk Institute in La Jolla California...This post discusses how a nationwide injunction against the FLSA is affecting universities' decisions to...

...incorporated into newly synthesized proteins instead of methionine. The labeled proteins can be detected using click-chemistry...turnover of individual proteins in situ.  For the visualization of specific proteins in situ immunocytochemistry...audiences. Right now, she is a science instructor at The Innovation Institute in Boston. When Eugenia is not ...protein is decreased in Alzheimer’s disease patient’s brains. However, it is not known why or how this happens...degradation. These processes control the abundance of proteins in cells. Homeostatic regulation of protein abundance...type. However, we cannot distinguish new from old proteins within that pool. Therefore, it is not possible...used to facilitate the visualization of specific proteins in cells. Tagged protein can easily be overexpressed...

...interest. Tagged proteins are typically expressed at much higher levels than endogenous proteins. Data obtained...Immunoprecipitation (IP) uses immobilized antibodies, or immunoglobulins, to isolate a specific protein out of a complex...study how a target protein interacts with other proteins or nucleic acids. Read on to learn more about ...optimized strategies for differing amounts of target proteins or antibody bonding strengths.    . Steps...from cells or tissues contain a complex mix of proteins seen as green squares, purple circles and blue...collected by centrifugation or a magnet. 4) Unbound proteins are washed away. 5) The target protein is eluted...eluted.     In the first method, an antibody against a target protein is immobilized, or tethered, on...

...interactions as well as be used for both soluble and insoluble proteins, which can be a challenge for classic antibody...can take hours to label proteins and APEX2, although it can rapidly label proteins, requires H2O2 which ...interacting proteins in one simple experiment. PCA is an in vivo technique where two proteins are fused ...interaction of two proteins while also biotin labeling and purifying additional proteins from a complex of... and an RNA component that contains an RNA motif of interest. Any proteins that bind the RNA motif will...interacting cell surface proteins with a biotin tagged LPETG peptide. Biotin tagged proteins can then be purified...There are approximately 2-4 million proteins per cubic micron in bacteria, yeast, and mammalian cells...

... the presence of proteins from samples containing a mixture of proteins. The proteins are separated based... Antibodies, also known as immunoglobulins, are ~150 kDa, Y-shaped proteins that are both a natural part...region. Two immunoglobulin (Ig) heavy chains and two Ig light chains make up each antibody molecule. The...the Ig heavy chains, and the rest of the Fab regions are completed with the Ig light chains. The variable...Flow cytometry is used to detect proteins on the surface of or inside whole cells. Antibodies - typically...antibodies are produced by B cells. They bind to proteins on the surface of extracellular pathogens such...such as parasites or microbes, or to proteins expressed on the surface of cells that have been infected ...

...Margaret Robinson’s lab, and more recently used by Stephen Royle’s lab, acutely depleted proteins from their...induction heterodimerization of proteins that contain rapamycin-binding domains, such as FKBP and FRB. The ...inactivation. Find the Robinson plasmids or the Royle plasmids and get to work! Robinson et al., Dev Cell...light-oxygen-voltage (LOV) sensing domains from algae into chimeric Opto-RTKs. The LOV domains bind flavin cofactors...phosphoserine (SP218) insertion was moderately enhanced while two phosphoserine insertions (SP218/SP222) was... to generate random mutagenesis libraries or to insert heterologous genes, reporters, or other markers...demonstrate that they can successfully create GFP fusion proteins with a variety of genes across the genome. Although...

...mix of proteins with a broad size range to be separated on a single gel.  Immobilizing proteins on a membrane...albumin or other proteins. The sticky areas on the membrane will bind to the proteins in the buffer, blocking...labeled. Instead, a conjugated species-specific secondary antibody is used to visualize the proteins. In this...Introduction to western blotting  In a western, proteins are: (1) separated by size, (2) transferred to...absence of a protein, but can also determine if proteins are being up- or downregulated in a system, detect... to standards, detect the cellular location of proteins, and can be a readout for protein interaction ...applied to the gel causes the negatively charged proteins to migrate toward the positive charge at the bottom...

...manner to Type I-F, but instead employ a Cas12k protein instead of three Cas proteins (Fig. 1A, bottom), and...transposase proteins (TnsA, TnsB, TnsC). The donor DNA (aka mini-transposon), which contains the DNA cargo...programmability: depending on the transposases involved, they insert either randomly, or at fixed genomic target sites...generation of tools for efficient and programmable insertions of large DNA cargos into bacterial genomes. This... This new capability was named INTEGRATE (Insertion of Transposable Elements by Guide RNA-Assisted Targeting...spacer sequence specifying the target site. Four proteins (TniQ, Cas8, Cas7, Cas6) that forms the QCascade...INTEGRATE. The requirement for multiple Cas proteins to form the Cascade targeting complex is a hallmark...

... prey proteins after screening. It is also possible to screen libraries of bait and prey against each ... expression of target proteins in heterologous systems (e.g. mammalian proteins in S. cerevisiae). ...activation and cell biology. During this period, proteins were successfully expressed as fusions that retain...domain (DBD) and transcriptional activation (TA) domains retain their individual activities when separated...could be combined effectively (3). In Figure 1 for instance, the DBD of the fusion protein shown is still ...also called the interaction trap), genes encoding proteins of interest(POI’s) are fused to TAs and DBDs (... then co-expressed in a yeast strain that also contains a reporter gene cloned downstream of a promoter...

...contain your insert of interest. Simply stated, ccdB makes cloning easier by selecting against vectors that...vector “backbone” in this system contains ccdB. A successful insertion will completely replace ccdB with...vectors in mind, any plasmid that contains the F plasmid (F’ strains) will also be resistant to CcdB, ...popular lab strains, such as NEB Stable, JM109, XL1 Blue or XL10 Gold, that are F’. These strains could possibly...that did not take up your insert. But exactly how does ccdB accomplish this? Let’s start with a brief ...antitoxin protein (CcdA) that protects the cell against the toxic CcdB. Cells that lose ccdA through the..., an investigator were to successfully clone an insert into the vector, the ccdB reading frame would be...

...for each insertion. Also, it remains challenging to screen for the genetically modified strains that contain...tagging proteins in a tissue specific manner, as well as 3-site destination vectors for inserting a tag ...tagging endogenous proteins with an auxin-inducible degron (AID) tag to generate proteins which can be rapidly...years ago, the growing spectrum of fluorescent proteins (FPs) has been an invaluable resource for studying...with FPs. Overexpression of fluorescently tagged proteins can lead to improper protein localization, protein...new user-defined DNA sequence (like GFP) can be inserted at the DSB. CRISPR has allowed scientists to tag...contain your newly tagged gene. Most insertions can only be detected by labor-intensive processes like...

...at allencell.org and the Allen Institute website at alleninstitute.org. A classic challenge in cell biology...at allencell.org and the Allen Institute website at alleninstitute.org. References 1. Roberts, Brock, ...contributed by the gene editing team at the Allen Institute for Cell Science. Learn more by visiting the Allen...tagging and visualization of specific, individual proteins under endogenous regulatory control. But even ...even with the advent of CRISPR/Cas9 technology, inserting large tags into a precise location in the genome...genomic tagging experiments. Browse the Allen Institute for Cell Science Plasmid Collection Fluorescent...C-terminal tag, as shown in the example above, we inserted a linker and GFP tag at the end of the last exon...

...neuronal synapses for instance. Also these tags need to be fused to scaffold proteins in order to be expressed... by red fluorescent proteins such as tdTomato or RFP or blue fluorescent proteins like Cyan. The issue...expressed proteins than standard tags and can be used to distinguish highly similar proteins. For example... neuronal populations in the brain. The Allen Institute and the Janelia Research Campus in the USA have...Neuronal tracers such as epitope tags and fluorescent proteins (FP) have also been important in mapping, monitoring...tags and even multimeric tags are frequently insufficient for detection when the POI is weakly expressed...expressed. As these epitopes tags have no intrinsic visual signal, they cannot be detected directly in live...

...pdDronpa domains to the sites flanking the DNA-binding cleft of Cas9, the fluorescent domains dimerize... Cas9 activity Anti-CRISPR proteins are a highly diverse group of proteins that are able to block CRISPR-Cas...The use of nickases, prime editing, anti-CRISPR proteins, and other techniques all aim to improve targeting...uses genetically encoded tools, such as microbial opsins, to control cellular activities using light. In... calcium and integrin-binding protein 1 (CIB1) proteins. The goal of both groups was to create a system...developed by the Sata lab is composed of two fusion proteins: 1) the genomic anchor - an inactive, dead Cas9...Once triggered by blue light, the CRY2 and CIB1 proteins heterodimerize and move the activator into position...

... photoconvertible proteins are PA-GFP, Dendra2, and the mEOS proteins. Some proteins are reversibly switchable...you really know how fluorescent proteins glow?   Fluorescent Proteins (FPs) were first discovered over...categories of fluorescent proteins Photoactivatable / Photoconvertible: These proteins can switch their color... less).  For LSS proteins, the Stokes shift is ≥ 100nm. Specifically, these proteins are excited by UV...when labeling proteins that interact. Indeed, FRET is often used to determine if two proteins interact. ...wavelength than the exciting wavelength. Thus, FPs are proteins with this unique capacity. Many of these FPs are...thus tracking the localization of that biomolecule inside living cells. FPs classified by the emission color...

...model organism, they insert into the host’s genome. Then, the transposon insertion sites are detected with...transposase proteins (TnsA, TnsB, TnsC), and 4) the donor DNA or mini-transposon that contains the DNA cargo...and result in human disease. While transposon insertions can be detrimental, it also drives genomic evolution...that moves around. This DNA sequence encodes the proteins the transposon needs for transposition. Transposition...transposition.  Target site: Different transposons insert at different DNA sequences or target sites. Integration...duplication of this target site sequence at the site of insertion.  Types of transposons Transposable elements ... reverse transcribe back into DNA before being inserted into new target sites. This is similar to how ...

...fluorescent proteins that are photostable. On top of photobleaching, fluorescent proteins may display ...performed with soluble fluorescent proteins or localized fusion proteins and do not require dedicated plasmids...performed on purified fluorescent proteins. To avoid diffusion, the proteins are (i) trapped in microdroplets...course of the experiment. The best fluorescent proteins for live cell imaging can be excited many times...fluorescent protein. This can differ between fluorescent proteins in the same spectral class (Cranfill et al, 2016...confocal laser scanning microscopy, fluorescent proteins are excited with very intense light (for a brief...compare the photostability of different fluorescent proteins? Measuring photostability Since it is clear that...

...   Dye stains Dye stains are non-antibody-based stains that bind DNA or free amines of proteins. Depending...cytosol/nucleus, or visualizing DNA instead of proteins. Some stains can diffuse through the cell membrane...transmembrane proteins, such as receptors, or post-translational modifications on those proteins, like glycosylation...kits allow for labeling of cytosolic proteins, as well as proteins of the secretory pathway. Those can ...buffer to label intranuclear proteins of interest, you can label cytosolic proteins alongside the intranuclear...like many of the cluster of differentiation (CD) proteins found on immune cells. However, there are more...methods can include labeling markers with multiple proteins/antibodies, targeting markers found within the...

...phosphoserine-incorporated proteins The generation of pSer-incorporated proteins requires a molecular toolkit... Sep-OTS, contains the components required for the translation of pSer-incorporated proteins on a single...Niemi, a postdoctoral fellow at the Morgridge Institute for Research in Madison, Wisconsin. It is commonly...cited that approximately one-third of cellular proteins are modified through phosphorylation (1). However...of a protein for characterization. However, phosphoproteins are often low abundance, and thus difficult...incorporation of phosphoserine directly into recombinant proteins (5). Optimization of this system defined three...effectively incorporate phosphoserine into recombinant proteins, such as the kinase MEK1 (5). MEK1 is inactive...

...generate two more OptoNBs, one against mCherry and one against GFP. Insertion of AsLOV2 into loop 1 always...how AsLOV2 insertion into mCherry or GFP nanobodies affected binding of their target proteins. The surface-exposed...opposite effects: insertion into loop 1 caused dark-induced binding while loop 6 insertion caused blue-light... you wash several times to remove non-specific proteins, and then elute YFP.  Option #2: Opto-Nanobodies... purification You skip adding a tag to YFP and instead use OptoNBs. You fill a column with OptoNB coated... the lights, OptoNBs bind YFP and non-specific proteins flow through. To elute YFP, you turn on the blue...ability of its nanobody partner, so it’s important to insert AsLOV2 at a spot in the nanobody that preserves...

...vector that contains everything needed to replicate inside a host cell but with space to insert your genetic...highly-specific recombinant antibodies against three of the six human glypican proteins (GPC1, GPC3, and GPC4). All...mini counterpart label nearby proteins with biotin. The biotinylated proteins can then be isolated and identified...validated viral packaging and strong expression after inserting EGFP and provide step-by-step protocols for viral...our bacterial plasmid set for expressing tagged proteins, and stay tuned for mammalian expression and AAV...and therapeutic potential. Our partners at the Institute for Protein Innovation (IPI) recently released...with the pithy name "site-specific target-primed insertion through targeted CRISPR homing of retroelements...

...scientists have used fluorescent proteins to observe what’s happening inside cells for at least a couple of...earlier described. “Until our proteins, there were lots of fluorescent proteins made from jellyfishes and ...decades. Green is the classic color, but fluorescent proteins are available in a variety of hues. While those... applications, Verkhusha and his lab at Albert Einstein College of Medicine in New York recognized their... The key to their bacterial phytochome-derived proteins, iRFP670, iRFP682, iRFP702, iRFP713 and iRFP720... The challenge then was to develop fluorescent proteins that would fall within that near-infrared range... and that’s exactly what iRFPs achieved. Those proteins allowed a signal-to-background ratio in mammalian...

...bacterial genome, and inserted it into E. coli to express and then test for any proteins that could cut RNA...the lab instead turned to phage serine integrases, where they saw 10-20% integration of inserts up to 36...Schematic of programmable gene insertion with PASTE. The PASTEsystem involves insertion of landing sites via Cas9...single-stranded nicks instead of the double-stranded breaks used in other insertion systems. Off-target ...nonetheless a new subtype. Cas7-11 had combined all the domains into a single protein, shedding the complexity ...Cas7-11   Use The Cas7-11 targeting system contains two pieces: a single protein, computationally ...computationally described as a fusion of four Cas7 proteins and a putative Cas11-like protein, and a guide RNA. The ...

..., the red fluorescent proteins form tetramers. In general, fluorescent proteins have a natural affinity...true monomeric fluorescent proteins. Concluding remarks Many fluorescent proteins are available, and their...aggregation The tendency of purified fluorescent proteins to form homodimers or higher order oligomers can...-PAGE under non-denaturing conditions separate proteins based on size. Whether higher molecular weight...protein, reflect the likelihood that fluorescent proteins interact or oligomerize in cells? This question...question is largely unanswered. Ideally, fluorescent proteins should act as inert modules that fail to homodimerize...homodimerize in cells. To understand how well fluorescent proteins approach this ideal situation, cellular assays...

...Rep78 and Rep68 contains a DNA-binding and endonuclease domains, all four Rep proteins share a common ...which contains the AAV structural and packaging genes, the adenoviral helper plasmid which contains the ...the proteins needed for the virus to replicate,  and the transfer plasmid which contains the viral genome...Rep proteins. Alternative splicing generates the different C-terminal regions of the Rep proteins. While...common helicase and ATPase domains. To function, the Rep proteins form oligomers of Rep 78/68 or Rep52/40. ... start sites generate the three VP proteins. The three VP proteins assemble into capsid at a 1:1:10 ratio...gene of interest can be packaged into the virus instead.  Next let’s take a closer look at the rep and ...