RecA-independent recombination between direct repeats of IS50

Zupancic, T.J.; Marvo, Sandra L.; Chung, Jae Hoon; Peralta, Ernest G.; Jaskunas, S R

doi:10.1016/0092-8674(83)90444-0

Cited by 12 publications

(13 citation statements)

References 20 publications

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“…If such cointegrates are intermediates in TnS transposition, then resolution may be partly dependent on host recombination/repair functions (e.g., type RecBC RecF nucleases) that may vary between bacteria. Previous results (26) suggest that IS50 p58 protein can catalyze not only transposition but also resolution of tandem;, directly repeated IS50 sequences that configure transient cointegrate structures in E. coli. However, other studies have concluded that TnS transposes in E. coli by a conservative, post-replicative mechanism because cointegrate structures are not observed (1, 2).…”

Section: Discussionmentioning

confidence: 95%

“…Because KpnI digestion of mutant genomic cointegrates would be expected to yield recombinant plasmids containing an IS50 triplication, the stability of such recombinant plasmids was suspect on theoretical grounds. At least two copies of IS50 would be in a directly repeated orientation and thus be a template for ISSOR-mediated resolution (26) or homologous recombination, both of which would result in excision and loss due to segregation of the recombinant DNA insert. Therefore, in a second protocol, EcoRI was substituted for KpnI; EcoRI cuts the cat gene DNA sequence of pVP2021 once (Fig.…”

mentioning

confidence: 99%

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Vector insertion mutagenesis of Rhizobium sp. strain ORS571: direct cloning of mutagenized DNA sequences

Donald¹,

Raymond²,

Ludwig³

1985

J Bacteriol

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When the limited-host-range plasmid pVP2021 carrying Tn5 was mobilized into Rhizobium sp. strain ORS571 and stable acquisition of TnS was selected, ORS571 plasmid-genome cointegrates were exclusively obtained; direct TnS transposition was never observed. In every case, genomic cointegrates exhibited an additional (third) IS50 element that bordered VP2021 DNA sequences but maintained a single Tn5 element. Genomic cointegrates containing IS50 triplications were stable; neither phenotypic reversion nor resolution was detectable. Auxotrophic mutant strains (vector insertion mutants) were identified at expected frequencies among derivatives carrying ostensibly random genomic pVP2021 insertions; N2 fixation (Nif)-defective vector insertion mutants were observed among these derivatives at a frequency of 10-3. The presence of integrated pVP2021 in ORS571 nif::VP2021 mutant genomes enabled VP2021 to constitute an endogenous cloning vector. After EcoRI or KpnI digestions, genomic nif::pVP2021 DNA sequences contiguous with integrated pVP2021 were directly cloned as new replicons without addition of an exogenous vector. Recombinant plasmids derived from two such nif::pVP2021 mutants hybridized to previously analyzed ORS571 Nif DNA sequences. Recombinant plasmid DNA and ORS571 Nif region DNA were found to be colinear; pVP2021 insertions could be accurately mapped. pVP2021 insertion-mutagenesis thus allows the direct cloning of ORS571 gene sequences for which mutant phenotypes can be selected or screened.

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Section: Discussionmentioning

confidence: 95%

mentioning

confidence: 99%

Vector insertion mutagenesis of Rhizobium sp. strain ORS571: direct cloning of mutagenized DNA sequences

Donald¹,

Raymond²,

Ludwig³

1985

J Bacteriol

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show abstract

“…Either the loss of homologous recombination was very recent, or such transfers could have occurred in a RecA-independent manner. For example, plasmids could have been transferred into Lv-StB cells, followed by the RecA-independent transposition of genes to the chromosome (Harmer and Hall, 2016; Zupancic et al, 1983).…”

Section: Discussionmentioning

confidence: 99%

Horizontal gene transfer to a defensive symbiont with a reduced genome amongst a multipartite beetle microbiome

Waterworth¹,

Flórez

Rees³

et al. 2019

Preprint

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19The loss of functions required for independent life when living within a host gives rise to 20 reduced genomes in obligate bacterial symbionts. Although this phenomenon can be 21 explained by existing evolutionary models, its initiation is not well understood. Here, we 22 32 33 38 Kroiss et al., 2010). Such relationships exist on a continuous spectrum of dependency 39 and exclusivity from the perspective of both the host and symbiont. Symbionts generally 40 become obligate after a prolonged period of exclusive association with the host, and the 41 3 symbionts that become obligate tend to carry out highly important functions for the host 42 (Latorre and Manzano-Marín, 2017; Lo et al., 2016; McCutcheon and Moran, 2012). For 43 example, mitochondria and chloroplasts, organelles that are required for energy 44 production and carbon fixation in eukaryotic and plant cells, originated from 45 endosymbiotic capture of alphaproteobacteria and cyanobacteria ~1.2 Bya and ~900 46

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“…Further, the recombination between the direct repeats of IS1 on pMCR16 can be prevented by introducing the AT mutation. The IS-catalyzed recombination resembles the homologous recombination observed between direct repeats of IS50 (38)…”

Section: Transcription From Pl During Transformation Of Pmcr16mentioning

confidence: 92%

Recombination in recA cells between direct repeats of insertion element IS1

Braedt¹

1985

J Bacteriol

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The IS1 sequences that flank the Tn9 chloramphenicol acetyltransferase gene as direct repeats recombine after transformation into an Escherichia coli recA strain. The recombination requires the lambda pL promoter on the plasmid. A plasmid that contains mutant IS1 elements does not recombine. These results indicate that this recombination requires an IS1-specific gene product. The recombinational activity of IS1 may resolve transient cointegrates formed during the transposition of IS1. I discuss a possible role for the lambda pL promoter.

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RecA-independent recombination between direct repeats of IS50

Cited by 12 publications

References 20 publications

Vector insertion mutagenesis of Rhizobium sp. strain ORS571: direct cloning of mutagenized DNA sequences

Vector insertion mutagenesis of Rhizobium sp. strain ORS571: direct cloning of mutagenized DNA sequences

Horizontal gene transfer to a defensive symbiont with a reduced genome amongst a multipartite beetle microbiome

Recombination in recA cells between direct repeats of insertion element IS1

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