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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Science Ltd
    Molecular microbiology 48 (2003), S. 0 
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Previous works have suggested that some gene complexes encoding a restriction (R) enzyme and a cognate modification (M) enzyme may behave as selfish mobile genetic elements. RM gene complexes, which destroy ‘non-self’ elements marked by the absence of proper methylation, are often associated with mobile genetic elements and are involved in various genome rearrangements. Here, we found amplification of a restriction–modification gene complex. BamHI gene complex inserted into the Bacillus chromosome showed resistance to replacement by a homologous stretch of DNA. Some cells became transformed with the donor without losing BamHI. In most of these transformants, multiple copies of BamHI and the donor allele were arranged as tandem repeats. When a clone carrying one copy of each allele was propagated, extensive amplification of BamHI and the donor unit was observed in a manner dependent on restriction enzyme gene. This suggests that restriction cutting of the genome participates in the am­plification. Visualization by fluorescent in situ hybridization revealed that the amplification occurred in single cells in a burst-like fashion that is reminiscent of induction of provirus replication. The multiplication ability in a bacterium with natural capacity for DNA release, uptake and transformation will be discussed in relation to spreading of RM gene ­complexes.
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 191 (1983), S. 213-220 
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary From Escherichia coli cells we purified ‘8’-shaped dimeric molecules in which two circular DNA molecules of bacteriophage lambda were joined at a homologous site. Some of them had a complex junction which we interpreted as being two closely spaced Holliday structures because of (i) superhelicity of the molecule, (ii) the sedimentation rate of the molecule in sucrose gradients, and (iii) the appearance in the electron microscope. Other ‘figure-eights's’ had two separate homologous junctions, presumably two Holliday bridges. A possible role for these ‘double Holliday structures’ in UV-stimulated recA-dependent recombination in vivo is discussed.
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  • 3
    ISSN: 1617-4623
    Keywords: Homologous recombination ; recA ; recF ; Holliday structure ; Mismatch repair
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Gene conversion, the non-reciprocal transfer of sequence information between homologous DNA sequences, has been reported in lower eukaryotes, mammals and in Escherichia coli. In an E. coli rec + strain, we established a plasmid carrying two different deleted neo genes (neoDL and neoDR) in an inverted orientation and then selected for homologous recombination events that had reconstructed an intact neo + gene. We found some plasmids that had apparently experienced intramolecular gene conversion. Further evidence, however, suggests that they are products of multiple rounds of reciprocal crossing-over,apparently involving two plasmid molecules. First, most of the Neo+ clones contained multiple types of Neo+ plasmids, although the frequency of producing the neo + clones was low. Second, all the neo + clones also contained, as a minority, one particular form of dimer, which can be formed by reciprocal crossing-over between neoDL of one plasmid molecule and neoDR of another plasmid molecule. Third, in reconstruction experiments, we cloned and purified this dimer and transferred it back into the rec + cells. The dimer gave rise to clones containing multiple types of neo + recombinant monomers, including those apparent gene conversion types, and containing only few molecules of this dimer plasmid.
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  • 4
    ISSN: 1617-4623
    Keywords: Homologous recombination ; Plasmid linear multimer ; Yeast mating-type switching ; Antigenic variation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Gene conversion - apparently non-reciprocal transfer of sequence information between homologous DNA sequences - has been reported in various organisms. Frequent association of gene conversion with reciprocal exchange (crossing-over) of the flanking sequences in meiosis has formed the basis of the current view that gene conversion reflects events at the site of interaction during homologous recombination. In order to analyze mechanisms of gene conversion and homologous recombination in an Escherichia coli strain with an active RecF pathway (recBC sbcBC), we first established in cells of this strain a plasmid carrying two mutant neo genes, each deleted for a different gene segment, in inverted orientation. We then selected kanamycin-resistant plasmids that had reconstituted an intact neo + gene by homologous recombination. We found that all the neo + plasmids from these clones belonged to the gene-conversion type in the sense that they carried one neo + gene and retained one of the mutant neo genes. This apparent gene conversion was, however, only very rarely accompanied by apparent crossing-over of the flanking sequences. This is in contrast to the case in a rec + strain. or in a strain with an active RecE pathway (recBC sbcA). Our further analyses, especially comparisons with apparent gene conversion in the rec + strain, led us to propose a mechanism for this biased gene conversion. This “successive half crossing-over model” proposes that the elementary recombinational process is half crossing;-over in the sense that it generates only one recombinant DNA duplex molecule, and leaves one or two free end(s), out of two parental DNA duplexes. The resulting free end is, the model assumes, recombinogenic and frequently engages in a second round of half crossing-over with the recombinant duplex. The products resulting from such interaction involving two molecules of the plasmid would be classified as belonging to the gene-conversion type without crossing-over. We constructed a dimeric molecule that mimics the intermediate form hypothesized in this model and introduced it into cells. Biased gene conversion products were obtained in this reconstruction experiment. The half crossing-over mechanism can also explain formation of huge linear multimers of bacterial plasmids, the nature of transcribable recombination products in bacterial conjugation, chromosomal gene conversion not accompanied by flanking exchange (like that in yeast mating-type switching), and antigenic variation in microorganisms.
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 222 (1990), S. 185-191 
    ISSN: 1617-4623
    Keywords: Bovine papillomavirus type 1 ; Plasmid rescue ; Non-reciprocality ; Single clone analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Bovine papillomavirus (BPV) shuttle vectors replicate as a circular plasmid in mouse cell nuclei without impairing host cell viability. We used these vectors to analyze homologous recombination in mammalian cells. When several BPV-based plasmids carrying direct repeats were introduced into C127 cells, we detected many recombinant plasmid molecules that have lost the sequence between the repeats. Many recombinant type molecules as well as parental type molecules were detected in all the cell clones isolated for analysis. Sequencing after rescue of the plasmid inEscherichia coli showed that most of the recombinants were from accurate homologous recombination. When the repeats on the plasmid were in inverted orientation, no crossing-over type products were detected. We discuss possible mechanisms that explain these features.
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  • 6
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Genetic recombination of phage lambda DNA mediated by Rec function of Escherichia coli was studied in the absence of duplication, transcription, translation, and maturation. Cells were jointly infected with double amber mutants, λ D - F 1 - and λ S - R - , and incubated in the presence of chloramphenicol and rifampin. The am + recombinant DNA molecules formed within the cell were detected by in vitro packaging as viable recombinant phages. This system was used to measure the recombination activity of rec - bacteria. In recA or recA recB bacteria, the number of recombinant DNA molecules was about 1% of the rec + level. In contrast, almost normal numbers of recombinant DNA molecules were formed in recB or recC cells. Therefore, (1) the recombination mediated by recA function does not need de novo protein synthesis; all gene products required for the recombination are present in the cell. (2) It can occur without duplication, transcription, and maturation of recombining DNA molecules. (3) The ATP dependent DNase (exonuclease V) controlled by recB and recC genes is not required for formation of recombinant DNA molecules.
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  • 7
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The role of the recA gene product of Escherichia coli in genetic recombination was examined in a system where recombination takes place in the absence of protein synthesis. recA200 bacteria were infected with two mutant strains of phage lambda in the presence of chloramphenicol and rifampin, and the resulting recombinant DNA molecules were measured by in vitro packaging. When recA200 bacteria grown at a temperature that is permissive for RecA phenotype were transferred to a temperature that is restrictive for RecA phenotype in the presence of the inhibitors, recombination of the infecting phages was severely blocked. This result shows that the recombination activity of the recA200 cells is inactivated by the change of temperature even in the absence of protein synthesis. The most likely explanation of this result is that the recA protein is directly involved in the recombination detected in the presence of chloramphenicol and rifampin.
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  • 8
    Publication Date: 2017-02-01
    Print ISSN: 0038-0806
    Topics: Architecture, Civil Engineering, Surveying , Geosciences
    Published by Elsevier
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  • 9
    Publication Date: 2003-06-01
    Print ISSN: 0038-0806
    Topics: Architecture, Civil Engineering, Surveying , Geosciences
    Published by Elsevier
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  • 10
    Publication Date: 1998-09-01
    Print ISSN: 0168-9525
    Electronic ISSN: 1362-4555
    Topics: Biology
    Published by Cell Press
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