Glycine-induced cryotransformation of plasmids into Bacillus anthracis

Summary: Different cloning vectors (pC194, pBC16, pUB110, pBD10, pBD8, pAMβ1) and Bacillus anthracis plasmid pX02 were introduced into B. anthracis by a transformation method. To induce an artificial competence state for uptake of isolated plasmid DNA, the cultures were treated with glycine, to reduce cross-linking of peptidoglycan, followed by freezing and thawing. The procedure is extremely rapid and relatively efficient (maximum transformation efficiency about 103 c.f.u. per λg DNA) and allows different cloning vectors with molecular masses ranging from 1.8 to 17.7 MDa to be introduced into B. anthracis.