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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Insertional transposon mutagenesis in the Xylella fastidiosa citrus variegated chlorosis strain with transposome

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Author(s):
Koide, Tie [1] ; Silva Neto, José F. da ; Gomes, Suely Lopes ; Marques, Marilis V.
Total Authors: 4
Affiliation:
[1] Universidade de São Paulo (USP). Instituto de Química. Departamento de Bioquímica - Brasil
Total Affiliations: 4
Document type: Journal article
Source: Current Microbiology; v. 48, n. 4, p. 247-250, Apr. 2004.
Field of knowledge: Biological Sciences - Genetics
Abstract

To overcome the difficulty in obtaining mutants of the citrus strains of Xylella fastidiosa, we evaluated mutagenesis using the transposome system as a tool for the isolation of a large number of mutants. Electroporation of a commercial transposome system in X. fastidiosa CVC (Citrus Variegated Chlorosis) strain J1a12 yielded an efficiency of 1.2 × 103 kanamycin (Km)-resistant clones per g of DNA. Southern blot analysis demonstrated that the transposon was randomly inserted, and nucleotide sequence analysis indicated the presence of 9 bp direct repeats flanking the transposon insertion site. Analysis by PCR of one of the insertion mutants (clone J15) showed that the transposon was stable after eight passages in solid media. These results show that the transposome system can be used to generate a random mutant library of Xylella fastidiosa CVC strain. (AU)

FAPESP's process: 98/16251-0 - Establishment of conditions for the disruption genes potentially involved in Xylella fastidiosa pathogenesis
Grantee:Suely Lopes Gomes
Support Opportunities: Genome Research Grants