Patent Number: 6,251,676

Title: Mammalian cell two-hybrid system

Abstract: 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.

Inventors: Shioda; Toshihiro (Burlington, MA), Isselbacher; Kurt J. (Newton, MA)

Assignee: The General Hospital Corporation

International Classification: C12N 15/10 (20060101); C12Q 1/68 (20060101); C12N 015/63 (); C12N 005/00 (); C12N 015/00 (); C12Q 001/68 (); C12P 019/34 ()

Expiration Date: 06/26/2018