Method for introduction of genetic material into microorganisms and transformants obtained therewith

摘要

A method of introducing nucleic acid into microorganisms and microorganisms transformed by such method. Known methods for transforming microorganisms to provide them with new genetic information have limited applicability. Some microorganisms are difficult to transform or transfect by the known techniques. These microorganisms are called recalcitrants. Some lactic acid bacteria, especially Lactobacillus and Bifidobacterium species are difficult to transform by known techniques. The invention solves this problem by subjecting the microorganism to limited autolysis in or before the transformation process. Said limited autolysis is carried out by incubating the microorganism in low molarity electroporation buffer containing an osmotic stabilizer, generally at a pH between 4 and 8, at a temperature below 37 DEG C, such as between 0 and 10 DEG C. The invention provides criteria for selecting plasmids to transform the microorganism with, and suitable plasmids for transformation. Applications of the transformed microorganisms are also disclosed.