Membrane Protein Production in Insect Cells

Research output: Chapter in Book/Published conference outputChapter (peer-reviewed)peer-review

Abstract

Membrane proteins are an essential part of the machinery of life. They connect the interior and exterior of cells, play an important role in cell signaling and are responsible for the influx and efflux of nutrients and metabolites. For their structural and functional analysis high yields of correctly folded and modified protein are needed. Insect cells, such as Sf9 cells, have been one of the major expression hosts for eukaryotic membrane proteins in structural investigations during the last decade, as they are easier to handle than mammalian cells and provide more natural posttranslational modifications than microbial systems. Here we describe general techniques for establishing and maintaining insect cell cultures, the generation and amplification of recombinant baculovirus stocks using the flashBAC™ or Bac-to-Bac™ systems, membrane protein production, as well as the production of membrane preparations for extraction and purification experiments.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
EditorsI. Mus-Veteau
PublisherSpringer
Pages223-240
Number of pages18
Volume2507
ISBN (Electronic)978-1-0716-2368-8
ISBN (Print)978-1-0716-2367-1
DOIs
Publication statusPublished - 1 Jul 2022

Publication series

NameMethods in Molecular Biology
Volume2507
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Bibliographical note

Funding Information:
We are grateful for funding from the European Union’s Horizon 2020 research and innovation programme under Marie Sklodowska-Curie grant agreement No. 847419 (MemTrain) as well as the ERACoBioTech MeMBrane project and BBSRC (BB/R02152X/1) to A.D.G, A.J.R, and R.M.B.

Keywords

  • Animals
  • Baculoviridae/genetics
  • Genetic Vectors
  • Insecta/metabolism
  • Mammals/metabolism
  • Membrane Proteins/genetics
  • Protein Processing, Post-Translational
  • Recombinant Proteins/genetics
  • Sf9 Cells
  • Spodoptera/metabolism
  • Recombinant baculovirus
  • Sf9 insect cells
  • Membrane protein production

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