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Showing: 1 - 15 of 15 results
  1. Split-BioID: An Improved Method for Studying Protein-Protein Interactions

    Type
    Blog Post
    ...Split-BioID is BirA*, a promiscuous version of BirA, an E. coli biotin ligase. In BioID, BirA* is fused to...-BirA*) and C-terminal BiraA* (C-BirA*). These two fragments are fused to two proteins, here proteins ...occurs, BirA* activity is restored, and nearby proteins are labeled with biotin. For labeling, BirA* fusion...where BirA*, an E. coli biotin ligase is split into two non-functional fragments: N-terminal BirA* (N-BirA... interact, the two BirA* fragments complement each other to form a functional BirA* enzyme. When this ...within a ~10 nm radius of the bait. In Split-BioID, BirA* is first split into two non-functional fragments... other, they combine forces to form a functional BirA* enzyme. This type of experiment is called a protein...
  2. RaPID Detection of RNA-protein Interactions

    Type
    Blog Post
    ... method. RaPID borrows the E. coli biotin ligase BirA* from BioID and allows a researcher to identify ...labeling methods are not ideal for in vivo work. BirA* requires 16-18 hours to sufficiently biotinylate...methods rely on the promiscuous E. coli biotin ligase, BirA*, to biotinylate proteins that are within ~10 nm...components of RaPID. Ramanathan et al. also tweaked BirA* to increase its rate of biotinylation. The improved...improved enzyme, called BASU, is derived from a BirA* homolog found in B. subtilis. BASU was  compared head-to-head... head-to-head with the E. coli and BioID2 BirA*s in the context of RaPID for their ability to detect a...detectable signal at the same time point. E. coli’s BirA* was tested at its optimum 18 hour time point, but...
  3. Proximity Labeling: A Powerful Tool for Protein Complex Purification and Proteomic Mapping

    Type
    Blog Post
    ...compared to the standard E. coli BirA* mutant. RapID consists of this mutant BirA* and an RNA component that...2012). BioID utilizes the E. coli biotin ligase, BirA*, with a site mutation (R11G). This site mutation...scientists can simply express a bait protein fused to BirA* in cells and then incubate those cells with exogenous...beads or matrices. The bait protein can even target BirA* to specific subcellular locations, such as the ...35 kD TurboID, which has 15 mutations compared to BirA and the 28kD miniTurbo, which has the N-terminus...domain deleted and 13 mutations compared to wild type BirA. TurboID and miniTurbo both enable biotin labeling...close proximity. Split-BioID consists of a split BirA* molecule that can reassemble to restore activity...
  4. Plasmids 101: Biotinylation

    Type
    Blog Post
    ... and BirA Proteins can also be biotinylated by exploiting the specificity of the E.coli BPL, BirA. If ...biotinylating proteins with BirA. You can find plasmids that contain both AviTag and BirA at Addgene! Oligonucleotide...are biotinylated. In E.coli for example, the BPL, BirA, is responsible for biotinylating only one lysine...If you recall, BirA biotinylates only one lysine in the entire E.coli proteome. Using this knowledge, scientists...identified the 13 amino acid sequence required for BirA recognition and biotinylation. This 13 amino acid...added to either the N or C terminus of the protein. BirA has been shown to effectively biotinylate AviTag...
  5. In Vivo Biotinylation of Bacterial Fusion Proteins

    Type
    Blog Post
    ...achieved by inserting BirA into a pMal plasmid (NEB) thus creating the pMal-T-Avi-His/BirA plasmid now available...attempt to create a plasmid based on the biotin ligase BirA backbone (Eric Campeau, Addgene plasmid #26624) ...Octet, Pall ForteBio). Tips for using pMal-T-Avi-His/BirA to add biotin to your protein of interest Grow ...
  6. Multiplexed Capture of Promoter-enhancer 3D Chromatin Structures Using CRISPR

    Type
    Blog Post
    ...components: dCas9 with an added biotin acceptor site BirA, a biotin ligase a gRNA(s) targeting a single genomic...the loci of interest and dCas9 is biotinylated by BirA. From here, the CAPTURE method is similar to other...Original CAPTURE CAPTURE 2.0 Biotinylation enzyme BirA Endogenous biotin ligases Expression system Stable... line expressing 1) FLAG-biotin-tagged dCas9, 2) BirA, and 3) one or more gRNAs Two separate lentiviruses...
  7. Hot Plasmids - August 2020

    Type
    Blog Post
    ...modifying pMAL-T-Avi-His/BirA, a vector commonly used for in vivo biotinylation via BirA and AviTAG. pMAK contains...
  8. Fluorescent Protein Guide: Empty Backbones

    Type
    Collection
    ... Brightness pKa Maturation Structure Plasmids mKusabira-Orange (mKO) 548 559 31 5 4.5 hr Monomer pQC mKorange... mKorange IX - Mammalian Expression mKusabira Orange-C1 (mKO) - Mammalian Expression mKO-pBAD - Bacterial...
  9. CRISPR Guide

    Type
    Guide
    ... biotin acceptor site to dCas9 and co-expressing BirA biotin ligase, as seen in the CAPTURE system . The...
  10. CRISPR Guide

    Type
    Collection
    ... biotin acceptor site to dCas9 and co-expressing BirA biotin ligase, as seen in the CAPTURE system . The...
Showing: 1 - 15 of 15 results