...One of the limitations is the size of GFP: ~240 amino acids or about 28 kDa. GFP and its homologues have...multiple cloning site and therefore consist of random amino acid sequences or (ii) designed to form an inert...glycines, serines and other small, non-aliphatic amino acids. However, long linkers increase the chance.... To indicate the sequence, people often use terminology like “C-terminal fusion,” but I find this very...protein fusions, our experience is that a linker of 4 amino acids (PVAT, when we use the AgeI site in the multiple...

...associated with phosphorylation. However, these amino acids fail to recapitulate the true steric and charge-based... version of GFP will contain a phosphoserine at amino acid 17 instead of a glutamate which allows the ... of these expression vectors, the codon for the amino acid to be replaced with phosphoserine must be recoded... a phosphoserine) to control for effects of the amino acid substitution. Alternatively, you can express..."Cysteinyl‐tRNA formation: the last puzzle of aminoacyl‐tRNA synthesis." FEBS letters 462.3 (1999): 302...

...IMP organizers Rosario F. Godino, Rocío López-Diego, & Zafira Castano Corsino. When someone says “internationalization... developed by its Director, Dr. Zafira Castano Corsino, a junior faculty member at Harvard Medical School...International Mentorship Program Zafira Castaño Corsino is a Junior Faculty at Harvard Medical School/BWH...zcastan or on linkedln.       Rosario Fernandez-Godino is a Postdoctoral Researcher at MEEI-Harvard Medical...

...listed as the amino acid change and not a nucleotide change. The proper way to denote an amino acid mutation...list the one letter abbreviation of the wild type amino acid immediately followed by its position (number...followed by the one letter abbreviation of the mutated amino acid currently at that position. In contrast, unmutated...

...letters of the DNA get translated into the twenty amino acids. Once scientists established that DNA encoded...meganucleases. The storyline involves the resurrection of dinosaurs by extracting the DNA from fossilized amber and...the insatiable appetite of the moviegoers for dinosaurs wreaking havoc in a theme park.  Released in 1997...ideal. For the early generation gene editors, new amino acid recognition sites had to be designed each time...the movie involves The Rock and his super giant albino gorilla fighting off a monstrous wolf and crocodile...

...Tsien’s lab in 1998 (Griffin et al., 1998), this 15 amino acid peptide contains a cluster of four cysteines...cysteines (CCXXCC, where X is any non-cysteine amino acid) that reacts with biarsenical dyes such as FlAsH (...on your protein of interest or using non-natural amino acids to directly incorporate a fluorophore in your...popular self-labeling tag systems Tag Size (amino acids) Ligand Commercial ligands available ...varies Various Few Complicated Non-natural amino acids 1 Various Many Complicated   ...

...Lercher, Spicer, & Davis, 2015). By using a novel aminoacyl tRNA synthetase/tRNA pair, a researcher can lure...the ribosome into inserting a fluorophore-tagged amino acid at a stop codon (often amber i.e. UAG) engineered...site. During translation, the novel tRNA/unnatural amino acid finds the UAG codon in the ribosome, resulting...Synthetase Expression Host pMAH-POLY Polyspecific aminoacyl-tRNA Synthetase Mammalian Cells pDULE-ABK pyrrolysyl-tRNA...pyrrolysyl-tRNA synthase for aliphatic diazirine amino acids E. coli and Mammalian Cells pEVOL-pAzF synthetase...

...are problematic because they require radiolabeled amino acids. The use of radioactive materials is highly...applies metabolic labeling using a noncanonical amino acid, azidohomoalanine (AHA). In the technique, ... labeling is known as fluorescence noncanonical amino acid tagging (FUNCAT, Dieterich et al. Nature Neuroscience...B., and L. Gabriel. "Inhibition by puromycin of amino acid incorporation into protein." Proceedings of...

...autophagy. Cells are cultured with isotope-labeled amino acids for several hours to several days, followed...followed by a short period of growth with unlabeled amino acids. This washout period removes radio-labeled ...ecliptic pHluorin (SEP). They are connected by a 9 amino acid linker that’s fused to the C-terminus of DsRed.T3...PubMed PMID: 18094608 2. Sargsyan, A., Cai, J., Fandino, L. B., Labasky, M. E., Forostyan, T., Colosimo...

...highly conserved glutamine-fructose-6-phosphate aminotransferase (glmS) gene in the recipient bacteria due ...plasmids, and lack the ability to synthesize diaminopimelic acid (DAP), a component of the cell wall. Once...disease (S. aureus, Klebsiella pneumoniae, and P. aeruginosa) in addition to Vicrio casei, a bacteria isolated...Enterobacter aerogenes, Klebsiella pneumoniae, and P. aeruginosa (all opportunistic pathogens) were grown in presence...

...catalyzes the conversion of a target ‘A’ to ‘I’ (inosine), which is treated as ‘G’ by cellular polymerases...converted to a G-C pair (Figure 1). As the cellular inosine excision repair is not as active as uracil excision... for rice crop improvement in the laboratory of Yinong Yang at PSU.   References 1. Komor, Alexis C., ...PMC5726555. 4. Xie, Kabin, Bastian Minkenberg, and Yinong Yang. "Boosting CRISPR/Cas9 multiplex editing capability...

...range of targets, including proteins, peptides, amino acids, drugs, metal ions, and cells (even pathogenic...aptamers hold a few important advantages over their amino acid-based counterparts, antibodies and enzymes,...structure stabilizes a 4-hydroxybenzylidene imidazolinone (HBI)-like compound allowing it to fluoresce...derivative 3,5-difluoro-4-hydroxybenzylidene imidazolinone (DFHBI) with relatively low fluorescence in ...

... oligonucleotides called morpholinos. When injected into zygotes, morpholinos complementary to a target...activity and does not work easily in zebrafish.) Morpholinos are a convenient and fast way to deactivate genes...whether a gene is deactivated via mutation or morpholino. This observation unlocked novel research into...

...identified the 13 amino acid sequence required for BirA recognition and biotinylation. This 13 amino acid sequence...processes such as gluconeogenesis, lipogenesis, amino acid metabolism and energy transduction. Biotinylation...Biotin’s exposed side chain can be cross-linked to amino acid functional groups in vivo via specialized chemicals...

...toxin genes isolated from E. coli or Psuedomonas aeruginosa. These toxins were a mix of Type VI secreted ...tested their expression under both rhamnose- and arabinose-inducible promoters. Tight control of toxin expression...though Khetrapal et al. also demonstrated that the arabinose promoter worked too. Rhamnose induction relies...et al. "Precision-engineering the Pseudomonas aeruginosa genome with two-step allelic exchange." Nature...

... that alter the nucleotide sequence but not the amino acid sequence of the translated protein.  These ... include the IPTG-inducible lac promoter, the arabinose-inducible pBAD promoter and the endogenous phage...communication and in vitro diagnostics.     References 1. Costantino N, Court DL. Enhanced levels of λ Red-mediated... 5. Thomason, L. C., Sawitzke, J. A., Li, X., Costantino, N. and Court, D. L. 2014. Recombineering: Genetic...

...Epitopes are between 5-8 amino acids long, with a typical length of 5 or 6 amino acids (Cruse et al., 2004...Antibodies may recognize a linearized sequence of amino acids, or only in their 3D conformation. Types of...and sensitivity, for example by changing certain amino acids to improve its stability. Additionally, the...

...construct, you clone all but one of your gRNAs into spectinomycin-resistant plasmids pX330S-2 to pX330S-(last ...systems use the Csy4 RNA nuclease from Pseudomonas aeruginosa. When overexpressed, Csy4 efficiently cleaves...and Cas9, which are separately inducible by L-arabinose and anhydrotetracycline (aTet), respectively. ...Addgene. 8. Xie, Kabin, Minkenberg, Bastian, & Yinong Yang. “Boosting CRISPR/Cas9 multiplex editing capability...

... from donors. Apart from fibroblasts, human keratinocytes from hair pluck, peripheral blood cells, and...Chondrocytes [27]         Adipocytes [28] Keratinocytes [29] Photoreceptors [30] Otic hair cells [...of induced pluripotent stem cells from human keratinocytes. Nat Biotechnol, 2008. 26(11): p. 1276-84. PubMed... human induced pluripotent stem cells into a keratinocyte lineage. Methods Mol Biol, 2014. 1195: p. 1-...

... is the nucleobase component of the nucleoside inosine. Created with BioRender.com. Although it was...To avoid confusion, we usually use different terminology when discussing base editors, like calling it...capable of chemically converting adenosine (A) to inosine (I, treated as G by the cell). Surprisingly, the...and Resources Glossary We introduced a lot of terminology in this post! Here’s a quick refresher if you...