서강대학교

초록 (요약문)

Protein-nucleic acid interaction is an important process in many biological phenomena. In this study, FAM modificated RNA-protein binding assay has been developed. A chloroethyl-containing amino acid was incorporated into a RNA-binding protein, and the mutant protein crosslinks with RNA labeled with fluorophore. The RNA-protein binding affinity was then measured with electrophoretic mobility shift assay (EMSA). In the Sex-lethal protein from Drosphila melanogaster, there are two RNA binding domain called RBD which binds specific to RNA. The RNA sequence selectivities of guanine-N7 alkylation produced by chloroethylating agent. FAM modificated RNA was crosslinked with chloroethoxy carbonyl-L-lysine incorporated Sxl protein and measured with EMSA. WT and mutant protein were compared by competition assay. Therefore, Protein-RNA interaction can be easily analyzed with this process. But it still needs to optimized condition of competition assay. Next plan is to find optimized condition. Furthermore, another unnatural amino acid with high electrophilic functional group such as epoxide can be used. With this system, any protein can be screening by fusion to RNA binding protein.