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http://dx.doi.org/10.5423/PPJ.2004.20.3.229

Insertional Transposon Mutagenesis of Xanthomonas oryzae pv. oryzae KXO85 by Electroporation  

Lee, Byoung-Moo (National Institute of Agricultural Biotechnology, Rural Development Administration)
Park, Young-Jin (National Institute of Agricultural Biotechnology, Rural Development Administration)
Park, Dong-Suk (National Institute of Agricultural Biotechnology, Rural Development Administration)
Kang, Hee-Wan (Graduate School of Bio and Information Technology, Hankyoung National University)
Lee, Gil-Bok (National Institute of Agricultural Biotechnology, Rural Development Administration)
Hahn, Jang-Ho (National Institute of Agricultural Biotechnology, Rural Development Administration)
Publication Information
The Plant Pathology Journal / v.20, no.3, 2004 , pp. 229-233 More about this Journal
Abstract
The bacterial leaf blight, which is caused by Xantho-monas oryzae pv. oryzae, is the most damaging and intractable disease of rice. To identify the genes involved in the virulence mechanism of transposon TnS 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 deter-mined 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.
Keywords
Mutagenesis; Transposon; Tn5; Xanthomonas oryzae pv. oryzae;
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