Expression Screening of Human Integral Membrane Proteins Using BacMam

Methods Mol Biol. 2021:2199:95-115. doi: 10.1007/978-1-0716-0892-0_6.

Abstract

This chapter describes the step-by-step methods employed by the Structural Genomics Consortium (SGC) for screening and producing proteins in the BacMam system. This eukaryotic expression system was selected and a screening process established in 2016 to enable production of highly challenging human integral membrane proteins (IMPs), which are a significant component of our target list. Here, we discuss our recently developed platform for identifying expression and monodispersity of IMPs from 3 mL of HEK293 cells.

Keywords: BacMam; Baculovirus; FSEC; Gel filtration; IMAC; Insect cells; Mammalian cells; Protein purification; SEC chromatography; Transduction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Gene Expression*
  • Genetic Vectors / genetics*
  • HEK293 Cells
  • Humans
  • Membrane Proteins* / biosynthesis
  • Membrane Proteins* / genetics
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / genetics

Substances

  • Membrane Proteins
  • Recombinant Proteins