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Detection and activity of insertion sequences in environmental strains of Burkholderia
Author(s) -
Miché Lucie,
Faure Denis,
Blot Michel,
CabanneGiuli Estelle,
Balandreau Jacques
Publication year - 2001
Publication title -
environmental microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.954
H-Index - 188
eISSN - 1462-2920
pISSN - 1462-2912
DOI - 10.1046/j.1462-2920.2001.00251.x
Subject(s) - biology , plasmid , burkholderia , polymerase chain reaction , insertion sequence , transposition (logic) , genetics , southern blot , genome , gene , microbiology and biotechnology , mobile genetic elements , bacteria , transposable element , linguistics , philosophy
The presence of two insertion sequences, IS 406 and IS 407 , was tested by polymerase chain reaction (PCR) amplification in 25 strains representing 15 Burkholderia species and the close relative Ralstonia pickettii . A total of 50% of the 25 strains contained at least one of the two insertion sequences (ISs) and a statistically significant correlation was found between the occurrences of IS 406 and IS 407 . Moreover, PCR–RFLP studies of the amplified fragments showed that IS 406 is largely conserved among all the strains tested, whereas IS 407 is rather polymorphic. Transposition activity was studied in Burkholderia vietnamiensis TVV75, using the pGBG1 target plasmid. This entrapping plasmid permitted the isolation and characterization of three active IS, able to activate the plasmid‐borne tetA gene after transposition. Sequencing permitted the identification of these mobile genetic elements as isoforms of IS 402 , IS 407 and IS 1416 . PCR amplification products provided IS probes, which were used to determine the copy‐numbers of IS 402 , IS 407 and IS 1416 in the genome of B. vietnamiensis TVV75, by Southern blotting. Copy numbers are 12, 3 and 11 respectively. To our knowledge, this is the first description of active insertion sequences in B. vietnamiensis.