Heterologous high-level gene expression in the photosynthetic bacterium Rhodobacter capsulatus

Methods Mol Biol. 2012:824:251-69. doi: 10.1007/978-1-61779-433-9_13.

Abstract

The functional expression of heterologous genes in standard hosts such as Escherichia coli is often hampered by various limitations including improper folding, incomplete targeting, and missassembly of the corresponding enzymes. This observation led to the development of numerous expression systems that are based on alternative, metabolic versatile hosts. One such organism is the Gram-negative phototrophic nonsulfur purple bacterium Rhodobacter capsulatus. During photosynthetic growth, R. capsulatus exhibits several unique properties including the formation of an intracytoplasmic membrane system as well as the synthesis of various metal-containing cofactors. These properties make R. capsulatus a promising expression host particularly suited for difficult-to-express proteins such as membrane proteins. In this chapter, we describe a novel R. capsulatus expression system and its application.

MeSH terms

  • Blotting, Western
  • Cell Culture Techniques / methods*
  • Cell Fractionation
  • Cell Membrane / metabolism*
  • Cloning, Molecular
  • Electrophoresis, Agar Gel
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli
  • Gene Expression Regulation, Bacterial / genetics
  • Gene Expression Regulation, Bacterial / physiology*
  • Genetic Vectors / genetics
  • Plasmids / genetics
  • Polymerase Chain Reaction
  • Rhodobacter capsulatus / cytology
  • Rhodobacter capsulatus / genetics
  • Rhodobacter capsulatus / metabolism*
  • Transformation, Bacterial