Genetic transformation of the phytopathogenic bacteria, Erwinia chrysanthemi.
Erwinia chrysanthemi is an enterobacterium whose phytopathogenicity is due to its pectinolytic and cellulolytic activities. The CaCl2 mediated transformation procedure was successfully applied to two E. chrysanthemi wild type strains. The highest efficiency of transformation of E. chrysanthemi with pBR322 was found using 0.1 M CaCl2, 0.1 M MgCl2 treated cells and a heat pulse at 30 degrees C for 6 min. This yielded about 600 transformants per microgram of pBR322 DNA and 2.3 X 10(-6) per viable cell. Plasmid stability after twenty generations was lower than in E. coli: only 40-60% of the cells retained the plasmid in the absence of selective pressure. Based on this result, cloning in E. chrysanthemi with pBR322 vectors should therefore be possible, making it a potential host for cloning any gene for biomedical or industrial purposes.
D002122:Calcium Chloride / Q000494:pharmacology; D004269:DNA, Bacterial / Q000032:analysis; D004587:Electrophoresis, Agar Gel; D004885:Erwinia / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D008274:Magnesium / Q000494:pharmacology; D015636:Magnesium Chloride; D010957:Plasmids; D014170:Transformation, Genetic / Q000187:drug effects
D004269:DNA, Bacterial; D015636:Magnesium Chloride; D008274:Magnesium; D002122:Calcium Chloride