Chemical and electroporated transformation of Edwardsiella ictaluri using three different plasmids.
作者： Riccardo Russo , Victor S. Panangala , Raphael R. Wood , Phillip H. Klesius
DOI: 10.1111/J.1574-6968.2009.01702.X
关键词:
摘要: Transfer of DNA by conjugation has been the method generally used for genetic manipulation Edwardsiella ictaluri because, previously, attempts to transform E. uptake naked have apparently failed. We report here successful transformation seven strains using electroporation and two different chemical procedures [conventional calcium chloride (CaCl(2)) 'one-step' (polyethylene glycol, dimethyl sulfoxide MgSO(4)) protocols]. Seven were transformed three plasmids [pZsGreen, pUC18 pET-30a(+)]. The highest efficiency was achieved (5.5+/-0.2 x 10(4) transformants ng(-1) plasmid DNA) than with CaCl(2) (8.1+/-6.1 10(-1) plasmid) 'one-step transformation' protocol (2.5+/-2.7 plasmid). An efficient required only 0.2 ng compared 200 one-step protocols. stably maintained in grown presence antibiotic 12 or more passages. results this study show that can be routinely molecular organism, is a quicker easier performed conjugation.
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