Optimized E. coli expression strain LOBSTR eliminates common contaminants from His-tag purification

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avisTidsskriftartikelForskningpeer review

  • Kasper Røjkjær Andersen
  • Nina C Leksa, Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA
  • Thomas U Schwartz, Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA

His-tag affinity purification is one of the most commonly used methods to purify recombinant proteins expressed in E. coli. One drawback of using the His-tag is the co-purification of contaminating histidine-rich E. coli proteins. We engineered a new E. coli expression strain, LOBSTR (low background strain), which eliminates the most abundant contaminants. LOBSTR is derived from the E. coli BL21(DE3) strain and carries genomically modified copies of arnA and slyD, whose protein products exhibit reduced affinities to Ni and Co resins, resulting in a much higher purity of the target protein. The use of LOBSTR enables the pursuit of challenging low-expressing protein targets by reducing background contamination with no additional purification steps, materials, or costs, and thus pushes the limits of standard His-tag purifications.

OriginalsprogEngelsk
TidsskriftProteins - Structure Function and Bioinformatics
Vol/bind81
Nummer11
Sider (fra-til)1857-1861
Antal sider5
ISSN0887-3585
DOI
StatusUdgivet - nov. 2013

Se relationer på Aarhus Universitet Citationsformater

ID: 84341409