Patent ID: 11473092

Abstract:
One objective of the present invention is to provide a novel simple and efficient transformation method for gram-positive bacteria, the transformation method being capable of introducing a large-sized DNA into a host DNA of the gram-positive bacteria without damage. In addition, another objective of the present invention is to provide a method in which desired DNA segments are accumulated in a chromosome of a recipient (recipient bacteria) to enable an artificially designed huge DNA to be constructed, and in which a transformed cell does not cause any problems in terms of controlling the natural environment. The present invention relates to a transformation method for gram-positive bacteria by conjugative transfer, characterized to use a helper plasmid having an origin of DNA transfer (oriT) region is inactivated. Preferably, the helper plasmid is a plasmid in which an oriT region is inactivated from pLS20cat. Furthermore, this transformation method is preferably a method in which donor bacteria have a helper plasmid having an origin of DNA transfer (oriT) region is inactivated and a chromosome DNA or plasmid incorporating an origin of DNA transfer (oriT) region, and transfer the same to the recipient bacteria.