Method for converting single-copy BAC vectors to conditional high-copy pBAC/oriHC vectors

摘要

A single-copy BAC vector (containing or lacking an insert) is converted in a host cell into a conditional high-copy BAC vector by introducing a conditional origin of replication into the single-copy BAC vector. The conditional ori is introduced by site-specific recombination between the SC BAC vector and a vector that contains the conditional ori. The host cell comprises a recombinase that recognizes a site-specific recombination site on both the BAC vector and the conditional ori vector. In the presence of the recombinase, the conditional ori-containing vector recombines into the BAC vector to produce a high-copy BAC vector that can be conditionally amplified by activating the conditional origin of replication on command.