Large vectors and methods for producing large vectors

Abstract

The development of a method for producing vectors of at least 16kb in size from bacterial cells involves the following sequential steps: a) Collect bacterial cells containing the vector, with a size of at least 16kb, containing the inductive replication sequence; b) Inoculate the bacterial cells containing this vector into a culture medium; c) Culture the bacterial cells in the culture medium; d) Add one or more of the aforementioned inductive replication sequence inducers to the culture medium when the bacterial culture medium reaches an optical density at 000nm (000nm) of at least 20; e) Continue culturing the bacterial cells in the culture medium. 0 g) Arbitrary separation of bacterial cells from the culture medium, and h) Recovery of plasmids from bacterial cells. The invention also proposes a vector of at least 16kb containing an induction replication initiation sequence for use in these methods.