...specificity and binding affinity, can be produced in E. coli, and are both soluble and easily engineered.  Monobodies...

...infections. The Cas3 enzymes target several strains of E. coli and lethally chop their genomes into many, many ...

... students to propagate the plasmids in either E. coli (ori and ampR) or S. cerevisiae (2µm ori and URA3...

...CRISPR libraries that target mouse, human, fly, E. coli, and T. gondii genes.  2) What genetic modification...

...transformation amplification protocol is the Chong Zhang E. coli Genome-wide Inhibition Library. DNA and cell amounts...

...programs to engineer 14 chromoprotein sequences into E. coli BioBrick plasmids (meffRed, eforRed, asPink, spisPink...

...thetaiotaomicron, Mycobacterium, Pseudomonas, and E. coli. However, many non-model bacterial species lack ...

...is to express and purify His-tagged Cas9 from E. coli using a plasmid such as pET-28b-Cas9-His from Alex...

...  Most Addgene plasmids are maintained in the E. coli lab strain DH5a, but others may require strains ...

...propagated in the endA-mutated NEB Stable strain of E. coli. In addition, plasmids are typically prepared using...

...Berkmen, M. (2012). SHuffle, a novel Escherichia coli protein expression strain capable of correctly folding...

...purification process simpler. Due to its high yields, E. coli is often used for the production of Fab fragments...

...fluorescence-activated cell sorting (FACS) of E. coli transformed with aptamer libraries, the Jaffrey ...nucleotide aptamer they called Broccoli. As a product of SELEX and FACS, Broccoli is inherently more stable...new RNA-based scaffolds to contain and stabilize Broccoli cassettes, thereby extending their use into mammalian...mammalian cells. They’ve also dimerized Broccoli to further enhance its biosensor capabilities (see Fig...et al. “In-gel imaging of RNA processing using broccoli reveals optimal aptamer expression strategies....Addgene.org Find the Spinach Plasmids at Addgene Find Broccoli Plasmids at Addgene View All Fluorescent Protein...

...Activity Learn more about protein tags Read about E. coli strains for protein expression Additional Resources...

...Nanobodies are easily expressed in the cytosol of E. coli, however the nanobody proteins tend to get stuck...inclusion body problem and makes nanobody production in E.coli as simple as following a DNA miniprep protocol!... were expressed in SHuffle T7 Express competent E.coli (NEB) using IPTG induction. To biotinylate the ...

...strep-tag and, more recently, OLLAS (Escherichia coli OmpF linker and mouse langerin) and Sun Tag (Viswanathan...

...beta-lactam and quinolone resistance genes in E. coli using plasmid and phagemid delivery systems. Phagemids...

...fusions for analyzing gene expression in Escherichia coli and yeast. Methods Enzymol. 1983;100:293-308. PubMed...

...Using Francisella Cpf1 (FnCpf1), they employed an E. coli plasmid depletion assay to discover FnCpf1’s PAM...

... High levels of gene expression Promoter from E. coli tryptophan operon Repressible Gets turned off with...

...construct can be easily expressed and purified from E. coli, retains important functional properties, and is...

...transforming a backbone phagemid vector into an E. coli strain that is deficient in dUTPase and uracil deglycosidase...

...PCR reaction, no ligation is required since the E. coli you transform your PCR products into will efficiently...

...post: Synthetic Photobiology: Optogenetics for E. coli. Schmidl, et al. ACS Synth Biol. 2014 Nov 21...aggregation propensity in four expression systems: E. coli, wheat germ, HeLa, and Leishmania (Gagoski, et al... replication increases plasmid copy number in E. coli. The authors used pXPG to study the human GM-CSF... F30-2xdBroccoli are available, as well as 5S RNA F30-2xdBroccoli fusion and pET28c-F30-Broccoli controls...mammalian cells with Broccoli Vegetable tags, including Spinach, Spinach2, and Broccoli, can benefit your...scaffold allows insertion of two dimeric Broccoli tags (2xdBroccoli) to further enhance fluorescence of the...controls. Don’t just eat more broccoli, use Broccoli tags in your experiments too! Filonov, et al. Chem...

... RSMs? In our study (1), we implement RSMs in E. coli with two plasmids (Figure 3 below): Input Plasmid...

...recombination persists even in recombinase deficient E. coli and under conditions used to minimize recombination...

...Engineering production of functional scFv antibody in E. coli by co-expressing the molecule chaperone Skp. Front...

...demonstrated in 2005 that proteins fused with E. coli dihydrofolate reductase (eDHFR) could be labeled...

...prime editors with RT domains derived from the E. coli Ec48 retron RT and the S. pombe Tf1 retrotransposon...

...Bacterial Expression Construct SpCas9 Inhibition (E. coli) Mammalian Expression Construct SpCas9 Inhibition...

... respectively. Both Red Broccoli and Orange Broccoli are derived from Broccoli, and share similar properties...properties to Broccoli despite binding to a different small molecule. Notably, like Broccoli, Red Broccoli and ...was described in the same study as Red Broccoli and Orange Broccoli, two more aptamers that bind DFHO and...collection. Green fluorescent aptamers Spinach and Broccoli and their derivatives are aptamers able to bind...Cas9 and track the localization of the complex. Broccoli, the other green fluorescent aptamer, was identified...brightest fluorescence (Filonov et al., 2014). Broccoli is twice as bright as Spinach2 when bound to DFHBI...useful tool for in vivo studies. Dimerization of Broccoli increases its fluorescence by about 1.8-fold, ...

...of the E.coli BPL, BirA. If you recall, BirA biotinylates only one lysine in the entire E.coli proteome...in the entire proteome that are biotinylated. In E.coli for example, the BPL, BirA, is responsible for ...

...began with the Broccoli aptamer system to visualize the gRNA in living cells. Broccoli aptamers are nucleic...

...PubMed PMID: 24882160. Creed, Meaghan, Vincent Jean Pascoli, and Christian Lüscher. "Refining deep brain stimulation... Glutamatergic synaptic plasticity in the mesocorticolimbic system in addiction. Front Cell Neurosci. ...

...small molecule, are often named after vegetables; Broccoli, Spinach, and Corn, for example. Learn more about...

...kit in the next two years. References 1. Martz, Colin A., et al. "Systematic identification of signaling...

... is related to a variety of food crops such as broccoli, cabbage and kale to name a few. Despite Arabidopsis...

...better signal in vivo as compared to imaging with broccoli aptamers or MS2 tags using the same number of ...

...Assembled plasmids can be directly transformed into E.coli for plating, screening and generation of sgRNA....

...insulin-like growth factor I from culture medium of E.coli. Bio/Technology. 1987, 5:379-382. Bornhorst JA ...

...experiment!     References and Resources References Ascoli CA, Aggeler B (2018) Overlooked benefits of using...

...website and online content are rich in resources. Colin White and Anthony Collmann of White Consulting frequently...

...components:  a deaminase (TadA, originally from E.coli, named TadA7.10 in ABE7.10)  a catalytically impaired...

...and scientific writing.     References 1. Martz, Colin A., et al. "Systematic identification of signaling...

... High specificityHigh sensitivity Longer protocolIncreased chance of cross-reactivityChallenging to develop...

...48(8): 512–20. PubMed PMID: 20506501. 6. Logie, Colin, and A Francis Stewart. 1995. “Ligand-Regulated ...

...serves as the code of life. In 1944, Oswald Avery, Colin MacLeod, and Maclyn McCarty’s experiment debunked...