Biolistic transformation of prokaryotes: factors that affect biolistic transformation of very small cells.
Five bacterial species were transformed using particle gun-technology. No pretreatment of cells was necessary. Physical conditions (helium pressure, target cell distance and gap distance) and biological conditions (cell growth phase, osmoticum concentration, and cell density) were optimized for biolistic transformation of Escherichia coli and these conditions were then used to successfully transform Agrobacterium tumefaciens, Erwinia amylovora, Erwinia stewartii and Pseudomonas syringae pv. syringae. Transformation rates for E. coli were 10(4) per plate per 0.8 micrograms DNA. Although transformation rates for the other species were low (less than 10(2) per plate per 0.8 micrograms DNA), successful transformation without optimization for each species tested suggests wide utility of biolistic transformation of prokaryotes. E. coli has proven to be a useful model system to determine the effects of relative humidity, particle size and particle coating on efficiency of biolistic transformation.
Smith
F D
FD
Department of Horticultural Sciences, Cornell University, Geneva 14456.
Harpending
P R
PR
Sanford
J C
JC
D016960:Agrobacterium tumefaciens / Q000235:genetics*; D003470:Culture Media; D004269:DNA, Bacterial / Q000235:genetics; D004885:Erwinia / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D006371:Helium; D006813:Humidity; D009994:Osmolar Concentration; D010316:Particle Size; D010957:Plasmids; D011549:Pseudomonas / Q000235:genetics*; D014169:Transformation, Bacterial; D014414:Tungsten
D003470:Culture Media; D004269:DNA, Bacterial; D006371:Helium; D014414:Tungsten