Cell Biology Methods to Study Recombinant Proteins in Seeds

Methods Mol Biol. 2022:2480:61-80. doi: 10.1007/978-1-0716-2241-4_4.

Abstract

Seeds are an attractive platform for the production of recombinant proteins because of their excellent storage properties and their well-developed endomembrane system, which allows accumulation of the product within specialized storage organelles. Due to the presence of these additional organelles and the resulting complexity of intracellular protein trafficking it is interesting to investigate the transport and storage of a recombinant protein within seed tissues, its interactions with endogenous reserve proteins and its impact on the ultrastructure of the endomembrane system. Possible approaches include sequential extraction procedures, subcellular fractionation and 2D as well as 3D electron microscopy techniques such as electron tomography (ET) and serial block face scanning electron microscopy (SBF-SEM), which are described and discussed in this chapter.

Keywords: Cereal seeds; Density gradient centrifugation; Electron tomography; Endomembrane system; Serial block face scanning electron microscopy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Electron Microscope Tomography* / methods
  • Imaging, Three-Dimensional / methods
  • Microscopy, Electron, Scanning
  • Protein Transport
  • Recombinant Proteins / genetics
  • Seeds* / genetics

Substances

  • Recombinant Proteins