Mammalian two-hybrid system

摘要

The invention provides a method of using a reporter gene that encodes a fluorescent polypeptide to indicate that an interaction has occurred between a bait and a prey protein in a mammalian cell. An advantage of using a fluorescent reporter polypeptide is that an interaction between a bait and prey in a mammalian cell can be readily detected, e.g., within 96 hours. In another method of the invention, a prey plasmid contains an Epstein-Barr virus origin of replication (ori-P). The OriP permits the prey plasmid to replicate episomally and indefinitely without damaging the mammalian cell or integrating into the genomic DNA of the mammalian cell. Since such a plasmid is maintained episomally in a circular form, it can be readily recovered from the mammalian cell.