Expression and purification of active protein kinases from wheat Germ extracts

Boglárka Sonkoly, Viola Bardóczy, Tamás Mészáros

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Citation (Scopus)

Abstract

In vitro functional studies of eukaryotic kinases are often constrained by the availability of pure and -enzymatically active kinase of interest. Though numerous proteins have been synthesized by cell-based systems, in vivo production of properly folded, eukaryotic proteins remains a challenging task. Current wheat-germ-based cell-free in vitro translation systems present a plausible alternative for protein synthesis since majority of eukaryotic proteins could be obtained in their native folded form with general protocols. The use of special in vitro translation vectors with ligation-independent cloning sites and cleavable affinity tags eliminates further bottlenecks of the protein producing procedure and makes this system a reasonable method for simultaneous generation of active kinases.

Original languageEnglish
Title of host publicationPlant Kinases
Subtitle of host publicationMethods and Protocols
EditorsNico Dissmeyer, Arp Schnittger
Pages55-63
Number of pages9
DOIs
Publication statusPublished - Oct 24 2011

Publication series

NameMethods in Molecular Biology
Volume779
ISSN (Print)1064-3745

Keywords

  • Active kinase
  • Affinity purification
  • In vitro translation
  • Ligation-independent cloning
  • TEV protease
  • Wheat germ

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Sonkoly, B., Bardóczy, V., & Mészáros, T. (2011). Expression and purification of active protein kinases from wheat Germ extracts. In N. Dissmeyer, & A. Schnittger (Eds.), Plant Kinases: Methods and Protocols (pp. 55-63). (Methods in Molecular Biology; Vol. 779). https://doi.org/10.1007/978-1-61779-264-9_3