Transposition of Tn4560 of Streptomyces fradiae in Mycobacterium smegmatis.

Abstract:

:Tn4560 (8.6 kb) was derived from Tn4556, a Tn3-like element from Streptomyces fradiae. It contains a viomycin resistance gene that has not been used previously for selection in mycobacteria. Tn4560, cloned in a Streptomyces plasmid, was introduced by electroporation into Mycobacterium smegmatis mc(2)155. Tn4560 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 AseI 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 Tn4560 delivery: up to 28% of the potential M. smegmatis transformants acquired a stable transposon insertion. The data indicated that Tn4560 may be useful for random mutagenesis of M. smegmatis.

journal_name

FEMS Microbiol Lett

authors

Bhatt A,Stewart GR,Kieser T

doi

10.1111/j.1574-6968.2002.tb11016.x

keywords:

subject

Has Abstract

pub_date

2002-01-10 00:00:00

pages

241-6

issue

2

eissn

0378-1097

issn

1574-6968

pii

S037810970100550X

journal_volume

206

pub_type

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