Transformation of Erwinia chrysanthemi by Escherichia coli plasmids DNA.

We have demonstrated in this study that CaCl2-MnCl2 treatment is effective in the preparation of competent cells of Erwinia chrysanthemi for transformation. Plasmids pMB9 and pBR322, which are of Escherichia coli origins, were transformed into E. chrysanthemi at frequencies of 1.5 X 10(-7) and 4.5 X 10(-7) per recipient, respectively. The frequencies were 60- to 80-fold lower than those in the well established rec- E. coli; however, the procedure is practically useful in transformation experiments. The plasmids were maintained rather stable in the cells if antibiotics were included in the media. When transformed by pECl and pEC6, which were chimeric plasmids consisting of pBR322 and an E. chrysanthemi chromosomal DNA fragment of 2.4 and 3.5 Mdal, respectively, the transformation frequencies of E. chrysanthemi were at the same range of that by pBR322. Neither colony morphology nor the polypectate degradation ability was changed after transformation by the plasmids. In conclusion, we have established a system of molecular cloning in E. chrysanthemi SR120A exploiting pBR322 as a vector.