Functional expression of plant membrane proteins in Lactococcus lactis.

Sylvain Boutigny (1) , Emeline Sautron (1) , Annie Frelet-Barrand (1) , Lucas Moyet (1) , Daniel Salvi (1) , Norbert Rolland (1) , Daphné Seigneurin-Berny (1)

1 LPCV - Laboratoire de physiologie cellulaire végétale

The study of most membrane proteins remains challenging due to their hydrophobicity and their low natural abundance in cells. Lactococcus lactis, a Gram-positive lactic bacterium, has been traditionally used in food fermentations and is nowadays widely used in biotechnology for large-scale production of heterologous proteins. This system has been successfully used for the production of prokaryotic and eukaryotic membrane proteins. The purpose of this chapter is to provide detailed protocols for (1) the expression of plant peripheral or intrinsic membrane proteins and then for (2) their solubilization, from Lactococcus membranes, for further purification steps and biochemical characterization.

Mots clés: bacteria; Lactococcus lacti; membrane protein; protein overexpression; Heterologous protein; Protocol

Domaines: Sciences du Vivant [q-bio]; Biochimie, Biologie Moléculaire

Sylvain Boutigny, Emeline Sautron, Annie Frelet-Barrand, Lucas Moyet, Daniel Salvi, et al.. Functional expression of plant membrane proteins in Lactococcus lactis.. Methods in molecular biology, 1258, Humana Press Inc., pp.147-65, 2015, Methods in Molecular Biology, 978-1-4939-2205-5 978-1-4939-2204-8. ⟨10.1007/978-1-4939-2205-5_8⟩. ⟨hal-01141092⟩