Cell-Free Synthesis of Plant Receptor Kinases

Methods Mol Biol. 2017:1621:37-46. doi: 10.1007/978-1-4939-7063-6_4.

Abstract

The wheat germ cell-free protein synthesis system has been used as a eukaryotic protein production system since it was first reported in 1964. Although initially the productivity of this system was not very high, it has now become one of the most versatile protein production systems, thanks to the enhancements made by several groups. In this chapter, we report a protein production method for plant receptor kinases using the wheat cell-free system. We describe a method for the preparation of a cell-free extract from wheat germ, the split-primer PCR method for preparation of transcription templates, and the bilayer cell-free protein synthesis method.

Keywords: Cell-free protein synthesis system; Wheat germ extract.

MeSH terms

  • Arabidopsis / enzymology
  • Arabidopsis / genetics*
  • Arabidopsis Proteins / biosynthesis
  • Arabidopsis Proteins / genetics*
  • Cell-Free System / metabolism*
  • DNA Primers / genetics
  • DNA Primers / metabolism
  • DNA, Complementary / genetics
  • DNA, Complementary / metabolism
  • Genetic Vectors / chemistry
  • Genetic Vectors / metabolism
  • Isoenzymes / biosynthesis
  • Isoenzymes / genetics
  • Plant Extracts / chemistry
  • Polymerase Chain Reaction / methods
  • Protein Biosynthesis*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Receptors, Cell Surface / biosynthesis
  • Receptors, Cell Surface / genetics*
  • Seeds / chemistry*
  • Transcription, Genetic
  • Triticum / chemistry*

Substances

  • Arabidopsis Proteins
  • DNA Primers
  • DNA, Complementary
  • Isoenzymes
  • PEPR1 protein, Arabidopsis
  • Plant Extracts
  • RNA, Messenger
  • Receptors, Cell Surface