Open AccessHigh‐quality mutant libraries of Xanthomonas oryzae pv. oryzae and X. campestris pv. campestris generated by an efficient transposon mutagenesis system
Author(s) -
Sun Qihong,
Wu Wei,
Qian Wei,
Hu Jun,
Fang Rongxiang,
He Chaozu
Publication year - 2003
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1016/s0378-1097(03)00583-4
Subject(s) - transposable element , xanthomonas campestris , xanthomonas oryzae pv. oryzae , transposon mutagenesis , biology , xanthomonas oryzae , transformation (genetics) , mutagenesis , genetics , transposition (logic) , genome , xanthomonas , xanthomonas campestris pv. campestris , genomic dna , southern blot , mutant , gene , linguistics , philosophy
A novel transposon mutagenesis system for the phytopathogenic bacteria Xanthomonas oryzae pv. oryzae ( Xoo ) and X. campestris pv. campestris ( Xcc ) was developed using a Tn 5 ‐based transposome. A highly efficient transformation up to 10 6 transformants per μg transposon DNA was obtained. Southern blot and thermal asymmetric interlaced polymerase chain reaction analyses of Tn 5 insertion sites suggested a random mode of transposition. The transposition was stable in the transformants for 20 subcultures. Eighteen thousand and 17 000 transformants for Xoo and Xcc , respectively, were generated, corresponding to 4 X ORF coverage of the genomes. The libraries will facilitate the identification of pathogenicity‐related genes as well as functional genomic analysis in Xoo and Xcc .