Plant Pathol J > Volume 20(3); 2004 > Article
The Plant Pathology Journal 2004;20(3):229-233.
DOI:    Published online September 30, 2004.
Insertional Transposon Mutagenesis of Xanthomonas oryzae pv. oryzae KXO85 by Electroporation
Byoung Moo Lee, Young Jin Park, Dong Suk Park, Hee Wan Kang, Gil Bok Lee, Jang Ho Hahn
The bacterial leaf blight, which is caused by Xanthomonas oryzae pv. oryzae, is the most damaging and intractable diseases of rice. To identify the genes involved in the virulence mechanism of transposon Tn5 complex, which possesses a linearized transposon and transposase, was successfully introduced into X. oryzae pv. oryzae by electroporation. The transposon mutants were selected and confirm the presence of transposition in X. oryzae pv. oryzae by the PCR amplification of transposon fragments and the Southern hybridization using these mutants. Furthermore, transposon insertion sites in the mutant bacterial chromosome were determined by direct genomic DNA sequencing using transposon-specific primers with ABI 3100 Genetic Analyzer. Efficiency of transposition was influenced mostly by the competence status of X. oryzae pv. oryzae cells and the conditions of electroporation. These results indicated that the insertion mutagenesis strategy could be applied to define function of uncharacterized genes in X. oryzae pv. oryzae.
Key Words: Mutagenesis, Transposon, Tn5, Xanthomonas oryzae pv. oryzae
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