Controllable alteration of cell genotype in bacterial cultures using an excision vector

Abstract

We have used recombinant DNA techniques to construct a derivative of phage lambda, called an excision vector, which retains only those functions necessary for conditional maintenance of lysogeny and integration/excision. The tyrA+ gene was cloned on this excision vector, integrated into the Escherichia coli chromosome, and stably maintained and expressed under permissive conditions. Upon shift to non-permissive conditions, the excision vector and its passenger gene were very efficiently excised from the chromosome and lost, leaving a culture of Tyr- bacteria. This illustrates a new class of conditional mutations in which the genotype changes in response to external stimuli.