Membrane proteins (MPs) play a crucial role in the life activities of humans and other species, which account for about 30% in all sequenced genomes. Drug targeting sites, cell-to-cell signaling, and detection of the environment mainly depends on specific membrane receptor proteins. MPs have significant theoretical and practical value in industrial, environmental, national defense and other research areas. Olfactory receptor (OR) protein is one kind of typical MPs, belonging to G protein-coupled receptor (GPCR) family. ORs regulate the organism to hunt for food, escape from risks and search for spouses. ORs are mainly distributed in the vertebrate nasal cavity or insect antennae. A series of reactions will be stimulated when the odorants bind with osmoceptors. The stimulus signal will be converted into an electrical signal. Eventually pass to the nervous system and to make the appropriate instructions. It is tough to extract MPs directly from tissues. Heterologous expression of MPs makes it difficult to integrate proteins into membranes, which brings great challenges to the study of structure and function. Cell-free protein synthesis system(CFPS) is an efficient strategy to express proteins in vitro on account of the open system and independence of cell lives. CFPS puts forward new ideas for synthesis and assemblage of MPs. It will be a major breakthrough for the study of structures and functions of MPs when the self-assembly and dimer formation can be accomplished in vitro. Here, we summarize latest progress in using CFPS to express ORs and other MPs.