Mini-Tn7 vectors as genetic tools for gene cloning at a single copy number in an industrially important and phytopathogenic bacteria, Xanthomonas spp.
Transposon mini-Tn7 vectors insert into the chromosome of several Gram-negative bacteria in a site-specific manner. Here, we showed the application of mini-Tn7 as single copy site-specific integration vector system for Xanthomonas campestris pv. campestris. The transposition of the mini-Tn7 into the bacterial genome was detected at a Tn7 attachment (attTn7) site located downstream of glmS1. Furthermore, using a newly constructed vector pBBR1FLP2 containing the flipase (FLP) recombinase for site-specific excision of the sequence between the FLP recombinase target (FRT) sites, and a sacB counter selection marker, an unmarked mini-Tn7 insertion mutant was created. Mini-Tn7 insertion did not affect bacterial virulence on the tested plant. The mini-Tn7 and FLP-FRT systems also work well in Xanthomonas oryzae pv. oryzae.
Jittawuttipoka
Thichakorn
T
Department of Biotechnology, Faculty of Science, Mahidol University, Bangkok, Thailand.
Buranajitpakorn
Sarinya
S
Fuangthong
Mayuree
M
Schweizer
Herbert P
HP
Vattanaviboon
Paiboon
P
Mongkolsuk
Skorn
S
D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors; D008967:Molecular Biology / Q000379:methods*; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional / Q000379:methods*; D017422:Sequence Analysis, DNA; D016959:Xanthomonas campestris / Q000235:genetics*
D004251:DNA Transposable Elements; D004269:DNA, Bacterial