About: Single-step affinity purification of recombinant proteins using a self-excising module from Neisseria meningitidis FrpC     Goto   Sponge   NotDistinct   Permalink

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Description
  • Anotace v českém jazyce Vyvinuli jsme samoštěpící kotvu umožňující jednokrokovou afinitní purifikaci rekombinantních proteinů. Sestává ze samoštěpícího modulu o velikosti 250 aminokyselinových zbytků, který byl izolován z proteinu FrpC bakterie Neisseria meningitidis a C-koncové afinitní kotvy. N-konec modulu je fúzován s C-koncem požadovaného proteinu. Po vazbě fúzního proteinu z buněčného lyzátu k afinitní matrici a odstranění kontaminujících proteinů je pomocí vápenatých iontů indukováno specifické štěpení mezi zbytky Asp-Pro spojujícími požadovaný protein a samoštěpící kotvu. To vede k uvolnění požadovaného proteinu nesoucího na svém C-konci pouze jeden aspartátový zbytek navíc, přičemž samoštěpící kotva zůstane zachycena na afinitní matrici (cs)
  • We developed a self-excising tag allowing single-step affinity chromatography purification of untagged recombinant proteins. It consists of a 250 residue-long self-processing module of Neisseria meningitidis FrpC protein with a C-terminal affinity tag. N-terminus of the module is fused to the C-terminus of a target protein of interest. Upon binding of the fusion protein to an affinity matrix from cell lysate and washing out of contaminating proteins, site-specific cleavage of the Asp-Pro bond linking the target protein to the self-excising tag is induced by calcium ions. This results in release of the target protein with only a single aspartic acid residue added at the C-terminus, while the self-excising affinity tag remains trapped on the affinity matrix
  • We developed a self-excising tag allowing single-step affinity chromatography purification of untagged recombinant proteins. It consists of a 250 residue-long self-processing module of Neisseria meningitidis FrpC protein with a C-terminal affinity tag. N-terminus of the module is fused to the C-terminus of a target protein of interest. Upon binding of the fusion protein to an affinity matrix from cell lysate and washing out of contaminating proteins, site-specific cleavage of the Asp-Pro bond linking the target protein to the self-excising tag is induced by calcium ions. This results in release of the target protein with only a single aspartic acid residue added at the C-terminus, while the self-excising affinity tag remains trapped on the affinity matrix (en)
Title
  • Single-step affinity purification of recombinant proteins using a self-excising module from Neisseria meningitidis FrpC
  • Jednokroková afinitní purifikace rekombinantních proteinů založená na samoštěpícím modulu proteinu FrpC bakterie Neisseria meningitidis (cs)
  • Single-step affinity purification of recombinant proteins using a self-excising module from Neisseria meningitidis FrpC (en)
skos:prefLabel
  • Single-step affinity purification of recombinant proteins using a self-excising module from Neisseria meningitidis FrpC
  • Jednokroková afinitní purifikace rekombinantních proteinů založená na samoštěpícím modulu proteinu FrpC bakterie Neisseria meningitidis (cs)
  • Single-step affinity purification of recombinant proteins using a self-excising module from Neisseria meningitidis FrpC (en)
skos:notation
  • RIV/61388971:_____/08:00313119!RIV09-AV0-61388971
http://linked.open...avai/riv/aktivita
http://linked.open...avai/riv/aktivity
  • P(GA310/06/0720), P(KAN200520702), Z(AV0Z50200510)
http://linked.open...iv/cisloPeriodika
  • 10
http://linked.open...vai/riv/dodaniDat
http://linked.open...aciTvurceVysledku
http://linked.open.../riv/druhVysledku
http://linked.open...iv/duvernostUdaju
http://linked.open...titaPredkladatele
http://linked.open...dnocenehoVysledku
  • 394988
http://linked.open...ai/riv/idVysledku
  • RIV/61388971:_____/08:00313119
http://linked.open...riv/jazykVysledku
http://linked.open.../riv/klicovaSlova
  • asp-pro bond; frpc; purification (en)
http://linked.open.../riv/klicoveSlovo
http://linked.open...odStatuVydavatele
  • US - Spojené státy americké
http://linked.open...ontrolniKodProRIV
  • [B5957E20BE44]
http://linked.open...i/riv/nazevZdroje
  • Protein Science
http://linked.open...in/vavai/riv/obor
http://linked.open...ichTvurcuVysledku
http://linked.open...cetTvurcuVysledku
http://linked.open...vavai/riv/projekt
http://linked.open...UplatneniVysledku
http://linked.open...v/svazekPeriodika
  • 17
http://linked.open...iv/tvurceVysledku
  • Novák, Petr
  • Osička, Radim
  • Šulc, Miroslav
  • Šebo, Peter
  • Sadílková, Lenka
  • Linhartová, Irena
http://linked.open...ain/vavai/riv/wos
  • 000259401900021
http://linked.open...n/vavai/riv/zamer
issn
  • 0961-8368
number of pages
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