Review of lactose-driven auto-induction expression of isotope-labelled proteins.

Autor: Crowley EL; Environmental and Life Sciences Graduate Program, Trent University, 1600 West Bank Drive, Peterborough, ON, K9J 0G2, Canada. Electronic address: erikacrowley@trentu.ca., Rafferty SP; Department of Chemistry, Trent University, 1600 West Bank Drive, Peterborough, ON, K9J 0G2, Canada. Electronic address: srafferty@trentu.ca.
Jazyk: angličtina
Zdroj: Protein expression and purification [Protein Expr Purif] 2019 May; Vol. 157, pp. 70-85. Date of Electronic Publication: 2019 Jan 30.
DOI: 10.1016/j.pep.2019.01.007
Abstrakt: NMR is an important method in the structural and functional characterization of proteins, but such experiments typically require isotopic labelling because of the low natural abundance of the nuclei of interest. Isotope-labelled protein for NMR experiments is typically obtained from IPTG-inducible bacterial expression systems in a minimal media that contains labelled carbon or nitrogen sources. Optimization of expression conditions is crucial yet challenging; large amounts of labelled protein are desired, yet protein yields are lower in minimal media, while the labelled precursors are expensive. Faced with these challenges there is a growing body of literature that apply innovative methods of induction to optimize the yield of isotope-labelled protein. A promising technique is lactose-driven auto-induction as it mitigates user intervention and can lead to higher protein yields. This review assesses the current advances and limitations surrounding the ability of researchers to isotope label proteins using auto-induction, and it identifies key components for optimization.
(Copyright © 2019 Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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