The use of an improved transposon mutagenesis system for DNA sequencing leads to the characterization of a new insertion sequence of Streptomyces lividans 66

Fischer, Joachim E.; Maier, Holger R.; Viell, P.; Altenbuchner, Josef

doi:10.1016/s0378-1119(96)00409-x

Cited by 27 publications

(14 citation statements)

References 33 publications

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“…Preliminary sequencing of copies a/b and c of the repeats that bordered the 20.2 kb amplification unit showed that they corresponded to the IS1372 element discovered by Fischer et al (1996). This implicates the involvement of a transposable element in the tandem amplification in the terminal region, an example of which has been reported by Eichenseer et al (1994).…”

Section: Note Added In Proofmentioning

confidence: 56%

Instability of artificially circularized chromosomes of Streptomyces lividans

Lin¹,

Chen²

1997

Molecular Microbiology

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SummaryThe chromosomes of Streptomyces species are linear molecules, containing long terminal inverted repeats and covalently bound terminal proteins. These chromosomes undergo spontaneous deletions of the terminal sequences at high frequencies and become circularized in several cases examined. Artificial circularization of the Streptomyces lividans chromosome was also achieved by targeted recombination in vivo, in which the terminal inverted repeats of the chromosome were connected by a kanamycin resistance gene (aphII ). Under kanamycin selection, the circularized chromosomes harboured tandem amplifications of a 20.2 kb sequence that included the aphII gene flanked by direct repeats and deletions nearby. On release from kanamycin selection, the aphII amplifications and the neighbouring sequences were deleted from the chromosomes, rendering all the cultures kanamycin sensitive. The chloramphenicol resistance gene, which was prone to deletion in wild-type S. lividans, became much more stable in the kanamycin-sensitive derivatives. These results indicate that the telomeres and/or certain terminal sequences may be involved in the structural instability of Streptomyces chromosomes.

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Section: Note Added In Proofmentioning

confidence: 56%

Instability of artificially circularized chromosomes of Streptomyces lividans

Lin¹,

Chen²

1997

Molecular Microbiology

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“…For transposon mutagenesis the strains E. coli JM108 F9 lacI proAB + [ : : Tn5491] (Fischer et al, 1996) and E. coli HB101 (Boyer & Rouland-Dussoix, 1969) were used. Replication and plasmid stability of SCP2-derived plasmids were tested in Streptomyces lividans TK64 (Hopwood et al, 1983).…”

Section: Methodsmentioning

confidence: 99%

“…The vector pJOE803 (Altenbuchner & Eichenseer, 1991) contains the thiostreptonresistance gene from Streptomyces azureus (Thompson et al, 1980) inserted into the NruI site of pIC20H (Marsh et al, 1984). The positive selection vector pJOE2114 used for transposon mutagenesis and deletion formation is a pIC20R derivative described by Fischer et al (1996). The cloning vector pDS100 contains a ThaI fragment with the lacZa-fragment of pUC19 inserted between the BamHI and HindIII sites of pACYC177.…”

Section: Methodsmentioning

confidence: 99%

“…For DNA manipulations, including DNA restriction, ligation, agarose gel electrophoresis, fragment isolation from gels, transformation of E. coli and plasmid isolation, standard methods were used as described in Sambrook et al (1989) and Kieser et al (2000). Nested deletions in SCP2 restriction fragments for subcloning and for DNA sequencing were generated by transposon mutagenesis using the Tn1721-derived Tn5491 as described by Fischer et al (1996). In principle, fragments of interest were inserted into pJOE2114 and the plasmids brought into JM108 F9lacI proAB…”

Section: Methodsmentioning

confidence: 99%

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Streptomyces coelicolor A3(2) plasmid SCP2*: deductions from the complete sequence

et al. 2003

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“…In this species, the extremities of the chromosome consist of 30 kb TIRs, which contain two transposable elements : Tn4811 and IS1372 (Lin et al, 1993 ;Fischer et al, 1996). A physical genetic map of a 1 Mb region spanning both chromosome ends has been assembled .…”

Section: Introductionmentioning

confidence: 99%

Genetic instability associated with insertion of IS6100 into one end of the Streptomyces lividans chromosome

1999

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Analysis of 548 recombinant strains of Streptomyces lividans carrying chromosomal insertions of IS6100 revealed that six mutants contained DNA amplifications. The amplifications differed in size but included IS6100 sequences. Hybridization with representative cosmid clones containing sequences from the unstable regions of the chromosome indicated that, in each mutant, DNA rearrangements affected just one of the chromosome ends. The amplifications were derived either from a region immediately proximal to the terminal inverted repeat (TIR) or further distal, from a previously characterized type I amplifiable unit of DNA. There was no evidence for extensive deletions accompanying the amplifications and chromosome linearity was maintained with, at least in five mutants, clear evidence for no loss of either TIR. The nature of the rearrangements provides evidence that insertions affecting the integrity of a chromosome end can contribute to genetic instability in Streptomyces.

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The use of an improved transposon mutagenesis system for DNA sequencing leads to the characterization of a new insertion sequence of Streptomyces lividans 66

Cited by 27 publications

References 33 publications

Instability of artificially circularized chromosomes of Streptomyces lividans

Instability of artificially circularized chromosomes of Streptomyces lividans

Streptomyces coelicolor A3(2) plasmid SCP2*: deductions from the complete sequence

Genetic instability associated with insertion of IS6100 into one end of the Streptomyces lividans chromosome

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