...the proteins cooperatively can separate the DNA strands. Created with BioRender.com. Once DnaA breaks...it recruits helicases to continue unwinding the strands. This creates two replication forks that move in...elongation phase, a DNA polymerase synthesizes a new DNA strand at each replication fork. Termination sequences...act as a feedback mechanism to reduce further transcription. Replication is also regulated by the balance... concentration is controlled at the level of translation. RepZ mRNA must fold into a “pseudoknot” held...intramolecular base pairs for the protein to be translated. An antisense RNA called Inc, also encoded by...prevents the mRNA from folding and blocks RepZ translation (Asano & Mizobuchi, 1998). This system keeps ...

... and inhibit its own transcription. As copy number increases, RepA transcription is repressed and replication...indirectly inhibit replication through inhibiting translation of replication machinery proteins, or directly... division. They don’t simply rely on chance (or random diffusion) to make this happen.  There are several...CBPs and NTPase to go around, the plasmids are randomly positioned, leading to plasmid loss (Diaz et al... if these do not provide clarity, a wet lab transformation experiment will ultimately inform whether your... Partition-associated incompatibility caused by random assortment of pure plasmid clusters. Molecular ...classification and identification based on replication and transfer systems and host taxonomy. Frontiers in Microbiology...

...fluor. Transcribing and translating a protein fluorophore also takes time - bypassing translation altogether... could be screened for their affinity to a wide range of non-nucleotide targets. These RNA molecules, ...The key to aptamers’ flexibility is their broad range of targets, including proteins, peptides, amino ...ions, and cells (even pathogenic bacteria). That range is possible because aptamers can either fold to ...biosensors. Aptabeacons Fluorescence resonance energy transfer (FRET) technology has been studied and used in...fluorescence-activated cell sorting (FACS) of E. coli transformed with aptamer libraries, the Jaffrey group selected...

...Hola! Que tal? My name is Brandon Solis, and I am a rising senior studying Sociology and Dance at Skidmore...interests are very clear, I had no idea how to translate them into real world careers. I figured this internship...a way of challenging what I am used to. Brandon Solis, marketing intern. Something that I struggle...world, I was worried about how my ideas would be translated through Zoom calls and Google docs. I pondered... do was create content that was unique but also brand friendly. I introduced the team to a new design ...program, Canva, and I was able to create cute and on-brand templates for these posts. Some of recent ... Brandon Solis...

...scientists, tech transfer offices, research foundations, and funding initiatives such as Fast Grants that support...worked closely with the legal partners and Tech Transfer Offices at universities to make COVID-19 research-related...United States, and many universities and Technology Transfer Offices were, at least, temporarily closing, these...Texas at Austin University of California, San Francisco  University of Pennsylvania Stanford University...them available to industry, please reach out at techtransfer@addgene.org and we will initiate a conversation...conversation with you and your technology transfer office to discuss the possibilities for expanded sharing. ...

... tissues are nearly transparent. As Verkhusha explained it to me, the transparency window of mammalian...tumor. They wanted to find something better. Transparency window That’s exactly what they reported doing...proteins that would fall within that near-infrared range, and that’s exactly what iRFPs achieved. Those proteins...corals, but all of them fluoresce outside the transparency window of mammalian tissues,” Verkhusha said...near-infrared fluorescent proteins within this transparency window, so now, we could see deeper.” Adding... proteins that operate within the 750 to 800 nm range, and his team is also looking in the direction of...

...is composed of single stranded DNA, which is particularly well suited for strand invasion, a key step ... are then packaged into AAV virions via transient transfection of 293T cells with helpers, and used to...non-sequence specific frameshift mutations through aberrant double strand break repair, it has proven more challenging...identical targeting vectors delivered through transfection of naked DNA. Other investigators who had been...AAV-based gene editing via the introduction of a double strand break by using a targeted nuclease. Asuri et al...

...emphatically deny that I have a magic formula to guarantee a reader the faculty job of his/her dreams. There...currency of scientific success is publications and grants. 2. Write succinct and easy to understand documents...years. You will discuss the focus of your first grant, how you will overcome certain obstacles, alternative...sentences) What will be the focus of your first grant? (1-2 sentences) Simply stated, what are your...It's not about you and how you will finally be granted the faculty position you have craved since grad...how YOU will fit into a department and bring in grants. What kind of colleague will you be? Do you understand...takes to be successful in running a lab, getting grants, and writing papers? This means that once you get...

...dopamine (DA), a well-known reward chemical and neurotransmitter. But aside from DA’s famous role as the “happy...standard method of measuring DA and other neurotransmitters in the brain has been in vivo microdialysis...performed with a probe surrounded by a semipermeable membrane, which is inserted into the region of the brain...passive diffusion of DA across the semipermeable membrane from the extracellular fluid into the perfused... to a threatening looming stimulus in vivo in transgenic zebrafish Report DA release in live mice over...developments in the detection and measurement of neurotransmitters in various species and while exhibiting complex...Dopamine-sensitive adenyl cyclase: possible role in synaptic transmission." Science 174.4016 (1971): 1346-1349. PubMed...

...bound to anything? Is it a transmembrane protein, or is it know to translocate or fold in certain conditions...as part of performing a specific function. A transmembrane protein that is being used as a marker for flow...cell, instead of when it is attached to a cell membrane.  Picking the Right Antibody  It can get even ...looking for a protein of interest while it’s in a transition state or specific configuration. So how do you...

...reverse transcribe their gRNA into DNA, which is then transcribed into mRNA before being translated into ...genome types that can be either double stranded (ds) or single stranded (ss) (ie, dsDNA, ssDNA, dsRNA, positive... of virus. RNA genomes can be transcribed into mRNA, directly translated into proteins, or, in the case...

...factors that could be transferred between cells. At the time, these extranuclear agents of heredity were...was proposed a few years later by Élie Jacob and François Wollman and became the widely adopted name for...-factors. Like F-factors, R-factors could be transferred between bacteria via cell-to-cell contact; however...pSC102, with EcoRI and selected for E. coli transformants that were resistant to both. When this proved...Chizuko Ogata, and Sachiko Sato. “EPISOME-MEDIATED TRANSFER OF DRUG RESISTANCE IN ENTEROBACTERIACEAE VIII:...PubMed Central PMCID: PMC314833. Datta, Naomi. “Transmissible Drug Resistance in an Epidemic Strain of Salmonella...

...field is a series of fluorescence resonance energy transfer (FRET)-based calcium biosensors. Perhaps most ...developed and published in 2010 for visualizing the transcription factor cAMP-responsive element-binding protein...really important and when? That is not known.” Transition Metals Maarten Merkx of Technische Universiteit...particularly for the intracellular imaging of transition metal ions, such as zinc, copper, and iron. “...the sensors involves plenty of trial and error. Transition metals also present challenges in that they are... way to make a FRET sensor with a large dynamic range,” he says. Merkx recently applied this “trick” to...red-shifted FRET sensor for zinc using variants of mOrange2 and mCherry as donor and acceptor domains. He ...

...contamination from uncut plasmid. 4. Transformation Transform your ligation reaction into your bacterial...perform, at minimum, two transformations after a ligation: 1. Control Transformation containing the ligation... how many colonies you should expect in the transformation due to background re-circularization and contamination...competent cells. For most standard cloning, you can transform 1-2μl of your ligation reaction into competent...ligation mixture with backbone alone; 2. Transformation containing ligation mixture with insert and backbone...conduct a positive control to ensure that your transformation worked. You should also verify that you are...

... seems straightforward: E. coli bacteria are transformed with the library DNA, they grow and replicate... Each Addgene library has its own depositor transformation and amplification protocol, but the overall...Maintain representation of the library during transformation First, use enough DNA and competent bacteria...and we use electroporation to ensure a good transformation efficiency. For a typical library, Addgene ...growing the cells on solid plates, where each transformant forms its own colony, uninfluenced by the growth...concern, but Addgene still recommends plating the transformation mixture on solid media. The colonies are then...that were amplified preferentially). You can also rank the plasmids by abundance, scaled to 1, to create...

...more detailed level, the Gal4/UAS system is a transcription activation system co-opted from yeast (1). It...workhorse to turn genes on or off. Gal4 is a transcriptional activator that binds to UAS enhancer sequences...sequences found in DNA. It then recruits transcription machinery to the site to induce gene expression. Thus...reads “#” as a “pound symbol”. References 1. A. H. Brand, N. Perrimon, Targeted gene expression as a means...15262411. 4. T. D. Southall, D. A. Elliott, A. H. Brand, The GAL4 System : A Versatile Toolkit for Gene ...Lai, T. Lee, Genetic mosaic with dual binary transcriptional systems in Drosophila., Nat. Neurosci. 9, 703...The Q system: a repressible binary system for transgene expression, lineage tracing, and mosaic analysis...

...responses are varied; one outcome is directed transcription from a given promoter. To harness the power ...the RR (OmpR) via phosphorylation, promoting transcription (see figure below). Subsequent exposure to red...response regulator via phosphorylation, promoting transcription of a GFP reporter. Subsequent exposure to red...activity persisted in the inactive state. The dynamic range, defined as the difference between the lowest and... reduced system leakiness and increased dynamic range to around 100-fold. These improvements will allow...Applications of these systems The large dynamic range of these systems ensures that they can be used with...types of proteins, including Cas9 and dCas9-transcription factors, for which low expression levels are...

...Wondering how you can develop more transferable skills? See Developing Transferable Skills During Science Training...of the University of British Columbia. When transitioning from an academic science career path to a non-academic...advantage over other candidates. Write about your transferable skills Some of the skills you developed might...research. For example, did you help put together a grant application? Are you familiar with the publishing...have presented in some public lectures? These transferable skills could be meaningful for your potential...Originally on track for a research career, she transitioned into a communications role instead after she...

...codon, which is then transfected into the desired cells. A hybrid mRNA transcript containing luciferase...luciferase, usually serves as an indicator of transcription within cells, where detection of the reporter...Higher Lower Background Lower Higher Post-translational Modification Not Required Required Photobleaching...regulatory elements, such as promoters, enhancers and untranslated regions, or the effect of mutations of these...as the most efficient means to disrupt the cell membrane and deliver the substrate; however, secreted luciferase...luciferase and the gene of interest permits translation of functional luciferase, until miRNA target and degrade...

.... elegans. These dsRNAs target specific mRNA transcripts for degradation. Despite how easy these experiments...often completely ineffective or results in low penetrant phenotypes that are hard to study. To overcome...target gene between two inverted T7 promoters. Transcription from each promoter progresses past the specific...podcast segment A fundamental approach to studying membranes, organelles, and other subcellular compartments...enzymes function across a much broader temperature range than BioID, which only functions optimally at 37...and C. elegans.   Find the TurboID plasmids here! Branon et al. Nat Biotechnol. 2018 PubMed PMID: 30125270...This tetramer cleaves chlorophenol red-b-D-galactopyranoside on the assay paper turning it purple, allowing...