Site-directed gene disruption in Xylella fastidiosa

• Author(s): Gomes, Suely; Koide, T.; Marques, M.; Neto, J.;
• Abstract: As a first approach to generate mutations by DNA insertion, we have developed a shuttle vector, called pSP3. which replicates both in Escherichia coli and in Xylella. Vector pSP3 was constructed by ligating to the E coli plasmid pBluescript, a kanamycin resistance gene under the control of the Xylella 16S rRNA promoter and the indigenous Xylella plasmid pXF1.3. Transformation experiments have shown that pSP3 replicates stably in Xylella. When a DNA fragment encompassing part of the Xylella xpsD gene was cloned into pSP3, specific integration of the construct into this gene was observed in 10% of the transformants, as early as after two passages of the culture, These results indicate that this vector can be used to generate site-specific gene disruption by homologous recombination in Xylella fastidiosa. (C) 2002 Published by Elsevier Science B.V. on behalf of the Federation of European Microbiological Societies.
• Publication Date: Apr 2002
• Journal: Fems Microbiology Letters