Research On Membrane Protein Expression With Cell Free Protein Synthesis System

Olfaction plays a crucial role in organisms feeding,mating,and selecting the host and many other life activities.Olfactory signal transduction will be activated when odor bound with olfactory receptor that is of great importance for olfaction.Olfactory receptor is a kind of membrane protein which is notoriously difficult to express,especially soluble expression.That makes it difficult to elucidate the structure and function of olfactory receptors.We focused on Drosophila Canton-S olfactory receptors Or83 b,Or7a and Or42 a to figure out the most effective CFPS to express membrane protein.We did some research on components,temperature and reaction time of CFPS,and finally established the platform of CFPS which is fit for membrane protein.Or83 b,Or7a and Or42 a can be expressed by this platform and the highest protein yield was 313?g/ml.We also tested the effects of detergent type on soluble expression,Brij35,Brij58,Triton100,Triton114,NP40,Tween20 and Tween80 included.We finally figured out the most effective detergent for soluble expression,and the highest soluble yield was 148?g/ml.