As is evident from the human immunodeficiency virus epidemic, there is no systematic method for producing a vaccine. Genetic immunization is a new approach to vaccine production that has many of the advantages of live/attenuated pathogens but no risk of infection. It involves introducing DNA encoding a pathogen protein into host cells and has shown promise in several disease models. Here we describe a new method for vaccine development, expression-library immunization, which makes use of the technique of genetic immunization and the fact that all the antigens of a pathogen are encoded in its DNA. An expression library of pathogen DNA is used to immunize a host thereby producing the effects of antigen presentation of a live vaccine without the risk. We show that even partial expression libraries made from the DNA of Mycoplasma pulmonis, a natural pathogen in rodents, provide protection against challenge from the pathogen. Expression library immunization may prove to be a general method for vaccination against any pathogen.