...The E. coli strain Most E. coli strains can be used to propagate plasmids, but endA- E. coli are best...started: Will the plasmid be used exclusively in E. coli? Gram negative bacteria in general? Both Gram negatives...-unnatural amino acid mutagenesis in Escherichia coli. Biochemistry. 2013 Mar 12;52(10):1828-37. doi: ...

...dcm– strain of E. coli such as JM110 and re-purifying it. These specialized E. coli strains have been ...enzymes can effectively cleave DNA. Common lab E. coli K12 strains such as DH5alpha contain 3 methylases...the other hand, should be derived from a dam+ E. coli strain and will therefore have methylated adenines...Discovery of DpnI Review describing dam-/dcm- E. coli Resources on the Addgene Blog Keep up with the...

...,170 Bikard Lab E. coli Genome-wide Inhibition Library v1 115927 Inhibition E. coli Bikard N/A 19 92,919...Bikard Lab EcoWG1: E. coli Genome-wide Inhibition Library v2 131625 Inhibition E. coli Bikard N/A ∼5 21,417...Zhang E. coli Genome-wide Inhibition Library 113134 — 113138 113146, 113147 Inhibition E. coli Chong Zhang...Other Species Human Mouse Fly Yeast T. gondii E. coli S. pneumoniae M. tuberculosis M. smegmatis Green...Protospacer flanking site (PFS) Library 79153 Knockout E. coli Zhang N/A N/A N/A - The protospacers contained in...

...expressing E. coli aided in ultrasound imaging of the mouse gastrointestinal tract. The E. coli strain Nissle...created two constructs optimized for expression in E. coli: acoustic gene 1 (arg1) and acoustic gene 2 (arg2...20 hours of growth following acoustic collapse E. coli re-express arg1 gas vesicles and restore their ultrasound...

...decided to add a bit of fun. He mixed together 10 E. coli strains producing different pigments, and tasked...ambient light. Expressing these chromoproteins in E. coli would make it easy to distinguish colonies from ...students transform them into different strains of E. coli,” says Westenberg. During the transformation experiment...

...cyanobacteria, synthetic inverters in E. coli, and biosensors in E. coli: Čelešnik, Helena, et al. "Biosafety...synthetic biology or protein purification studies in E. coli. 800 and 1900 Series If you’re expressing two genes... be used to precisely tune your polycistronic E. coli expression constructs. Some applications of plasmids...in the kit can be used to purify proteins from E. coli, Bacillus subtilis, or Pichia pastoris depending...aggregation propensity in four expression systems: E. coli, wheat germ, HeLa, and Leishmania (Gagoski, et al... clone your segments of choice into either an E. coli or a mammalian expression vector in a one-pot reaction...

...interaction detection (RaPID) method. RaPID borrows the E. coli biotin ligase BirA* from BioID and allows a researcher...interactions. Both methods rely on the promiscuous E. coli biotin ligase, BirA*, to biotinylate proteins that...subtilis. BASU was  compared head-to-head with the E. coli and BioID2 BirA*s in the context of RaPID for their...no detectable signal at the same time point. E. coli’s BirA* was tested at its optimum 18 hour time point... generated a >30-fold enrichment compared to E. coli’s BirA*. Faster labeling kinetics also means that...

...some E. coli strains have natural antibiotic resistances, so make sure your plasmid and E. coli strain ... makes our lives. Plasmid transformation into E. coli is a fairly inefficient process– just 1 out of 10,000...are compatible! Check out this list of common E. coli genotypes and their natural resistances.  Additional...

...highest levels of nonlinear acoustic contrast in E. coli. We deleted an unannotated hypothetical protein,... with several inducible expression systems in E. coli. The arabinose-inducible promoter pBAD and a low-copy...pBAD-bARGSer-AxeTxe, gas vesicle expression in E. coli is easy: simply grow cells in Lennox LB (10 g/L ...expression in vivo is also straightforward: after the E. coli have colonized tumors, inject L-arabinose intraperitoneally...).     Fig. 1: Expressing bARGSer in E. coli. (a) Plasmid diagram of pBAD-bARGSer-AxeTxe. (b)...imaging of tumor colonization by bARGSer-expressing E. coli. The GV-specific ultrasound signal inside the tumors...

...dsRNA-expressing E. coli that are mixed with liver paste. Prior to mixing with the liver paste, these E. coli are transformed...the  induction of T7 polymerase encoded in the E. coli genome (usually controlled by IPTG). For this IPTG-induced...expression to work, students have to use a commercial E. coli strain that is designated as DE3. For Phagocata ...eaters and refuse to eat liver paste mixed with E. coli. Thus, to get the dsRNA into Phagocata morgani and...

...Check the bacterial strain Differences between E. coli host strains can impact plasmid yield. Check the...is advisable to check out the genotype of your E. coli to ensure it is suitable for your application. We... We have previously reviewed many common lab E. coli strains on the blog, and also recommend this handy...OpenWetWare for a more extensive listing of common E. coli strains. Increase the volume Starting with a larger...to Verify Your Plasmid Learn about Common Lab E. coli Strains Additional Resources on Addgene.org Find...

...phosphorylated amino acid to the genetic code of E. coli (Park et al., Science 2011). Follow up work from...the parent protein. Using the recoded strain of E. coli ( 68306 ) with the optimized phosphoserine orthogonal...flexible codon in genomically recoded Escherichia coli permits programmable protein phosphorylation. Pirman...Enhanced phosphoserine insertion during Escherichia coli protein synthesis via partial UAG codon reassignment...described in: Expanding the genetic code of Escherichia coli with phosphoserine. Park HS, Hohn MJ, Umehara T,...lab published a new strain of recoded E.coli (rEcoli XpS 192872 ) and an optimized phosphoserine orthogonal...phosphoprotein expression in rEcoli XpS in the Methods section link below (Methods for iSPI, rEcoli XPS, pSerOTS-C1...

...Plasmids 101: E. coli strains for protein expression Plasmids 101: Common Lab E. coli Strains Why Bacteria...Bacteria? E. coli are commonly used to store and replicate plasmids of all types, but beyond that, researchers...researchers also use bacteria like E. coli and their relatively well understood biology to answer many interesting... an N-terminally His-tagged TALEN fused to an E. coli codon optimized FokI endonuclease domain. pET-Sangamo-His...of a C-terminally His-tagged TALEN fused to an E. coli codon optimized FokI endonuclease domain. Return...can be used to express a protein of interest in E. coli , B. subtilis and P. pastoris . All plasmids use...

...protocol) At Addgene, we use the Mix & Go! E. coli Transformation Kit and Buffer Set  from Zymo Research...available via Zymo Research. In brief, we grow our E. coli in LB to log phase, then wash and resuspend the ... protocol has worked well for us for multiple E. coli strains, including DH5alpha, XL1 Blue, TOP10, ccdB...

...response to changing pH, and they can rupture E. coli spheroplasts to release encapsulated proteins. Video...in a single operon and were first expressed in E. coli in the 1980s  (Kanabrocki et al 1986). Much later...showed that these fluorescent R bodies can rupture E. coli spheroplasts (cells which have had their cell walls...Absence of toxicity of R-body producing recombinant E. coli on paramecia. European Journal of Protistology 48...

...plasmids suitable for other model systems, including E. coli, plants, Drosophila, and zebrafish!   Why use multiplexed...Assembly or SLIC cloning methods. A BsaI-based E. coli multiplexing plasmid is available from the Koffas...supplied on a separate plasmid. Multiplexing in E. coli Koffas Lab CRISPathBrick Multiplex Plasmid: This...arrays A, B, C, AB, AC, BC, and ABC). Nielsen Lab E. coli CRMAGE Plasmids: The CRMAGE system is a fast, multiplexable...incubation. Kondo Lab multiplexed base editing in E. coli The Kondo lab expressed a cytidine deaminase fusion...LVA tag) to achieve specific point mutations in E. coli. gRNAs are delivered on a different plasmid and ...Mediated Multiplex Transcriptional Repression in E. coli.” ACS Synthetic Biology 4(9) (2015):987-1000. PubMed...

...looking at an example plasmid, let's review how E. coli normally replicate their chromosomal DNA. Chromosomal...positive and negative regulators. The genome of E. coli is about 4.6 million base pairs long and contains...re-activate DnaA. These regulatory mechanisms allow E. coli to control chromosomal replication and balance population...concentration of plasmid. Similar to replication in E. coli, regulation primarily occurs by controlling the ...controlled and has a low copy number. Like the E. coli chromosome, it has a single ori and requires the...regulate plasmid replication. For example, the plasmid ColIb-P9 encodes the protein RepZ to drive its own replication...An antisense RNA called Inc, also encoded by the ColIb-P9 plasmid, is complementary to part of the sequence...

...these broad host range plasmids all replicate in E. coli, which is the most commonly used bacteria in lab...Erwinia carotovora, E. chrysanthemi, Escherichia coli, Gluconacetobacter xylinus, Gluconobacter spp., ...herbicola, E. rubrifaciens, E. stewartii, Escherichia coli, Klebsiella spp., Legionella pneumophila, Methylophilus...Brucella spp., Caulobacter crescentus, Escherichia coli, Gluconacetobacter xylinus, Paracoccous denitrificans...promoter-based expression systems for Escherichia coli and Agrobacterium tumefaciens. 3 Biotech, 13(3),... ferrooxidans, Vibrio salmonicida, Yersinia enterocolitica, Xanthomonas campestris, X. maltophilia ...

...knockout E. coli strain to assay for functional decaffeination in cells. The ΔguaB E. coli are missing... CBB5 and has been tailored for expression in E. coli. This operon is under the transcriptional control...Janthinobacterium sp. Moreover, a caffeine “addicted” E. coli strain expressing this operon can serve as a living...

...glycosylated proteins that cannot be produced in  E. coli or yeast cells. The only problem with baculovirus...bacterial host cell is almost always the answer. E. coli is definitely one of the most popular hosts for ...expression may be required if your protein is toxic to E. coli. Of all the vectors, pET, under the control of the...Systems Recombinant protein expression in Escherichia coli: advances and challenges Production of recombinant...