Improved electroporation of Rhodococcus equi.
The condition of an electroporation method was re-evaluated for the introduction of foreign plasmid DNA into Rhodococcus equi. The method is based on an electroporation of the bacteria made competent by culturing in a broth containing glycine and by heat shock at 50 degrees C. Transformation of R. equi could be achieved with a chloramphenicol-resistant shuttle vector originating from Rhodococcus fascians at an efficiency of about 10(4) transformants/microgram DNA. The bacteria were also shown to become competent when they were incubated with a chemical transformation buffer prior to washing with an electroporation buffer.
Sekizaki
T
T
National Institute of Animal Health, Tsukuba, Ibaraki, Japan.
Tanoue
T
T
Osaki
M
M
Shimoji
Y
Y
Tsubaki
S
S
Takai
S
S
D003470:Culture Media; D018274:Electroporation / Q000379:methods; D018014:Gene Transfer Techniques; D005990:Glycerol / Q000494:pharmacology; D005998:Glycine / Q000494:pharmacology; D010957:Plasmids; D011136:Polysorbates / Q000494:pharmacology; D016929:Rhodococcus equi / Q000187:drug effects / Q000254:growth & development
D003470:Culture Media; D011136:Polysorbates; D005990:Glycerol; D005998:Glycine