Open AccessTransposition of Tn 4560 of Streptomyces fradiae in Mycobacterium smegmatis
Author(s) -
Bhatt Apoorva,
Stewart Graham R,
Kieser Tobias
Publication year - 2002
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 - 10.1111/j.1574-6968.2002.tb11016.x
Subject(s) - streptomyces fradiae , mycobacterium smegmatis , transposable element , biology , plasmid , genetics , insertion sequence , viomycin , transposition (logic) , transposon mutagenesis , streptomyces , mutagenesis , gene , microbiology and biotechnology , genome , mutant , actinomycetales , bacteria , mycobacterium tuberculosis , tuberculosis , medicine , linguistics , philosophy , pathology , streptomycin , antibiotics
Tn 4560 (8.6 kb) was derived from Tn 4556 , a Tn 3 ‐like element from Streptomyces fradiae . It contains a viomycin resistance gene that has not been used previously for selection in mycobacteria. Tn 4560 , cloned in a Streptomyces plasmid, was introduced by electroporation into Mycobacterium smegmatis mc 2 155. Tn 4560 transposed into the host genome: there was no obvious target sequence preference, and insertions were in or near several conserved open reading frames. The insertions were located far apart on different Ase I macrorestriction fragments. Unexpectedly, the transposon delivery plasmid, pUC1169, derived from the Streptomyces multicopy plasmid pIJ101, replicated partially in M. smegmatis , but was lost spontaneously during subculture. Replication of pUC1169 probably contributed to the relatively high efficiency of Tn 4560 delivery: up to 28% of the potential M. smegmatis transformants acquired a stable transposon insertion. The data indicated that Tn 4560 may be useful for random mutagenesis of M. smegmatis .