Cloning and Expression of Riboflavin Synthase Gene in Rice and Preparation of Polyclonal Antibody against Riboflavin Synthase

Riboflavin synthase(RS) catalyzes the committed step in biosynthesis of riboflavin,which plays an important role in organisms.Here,riboflavin synthase gene in rice(OsRS) was cloned and expressed in Escherichia coli with high efficiency.Since the product of expressed OsRS occurred in the form of inclusion body,the protein was dissolved with 6 mol/L guanidine hydrochloride solution and then was dialysed with 8 mol/L urea solution.The rabbit anti-OsRS polyclonal antibody was prepared.Furthermore,antigen-specific immune responses were determined by the Western blot with OsRS protein expressed in E.coli and total proteins from rice leaves as antigens.The results will be potential for studying catalytic activity and function of riboflavin synthase in rice.