原创力文档restriction and sequence alterations affect dna uptake sequence-dependent transformation in neisseria meningitidis限制和序列变化影响脑膜炎奈瑟菌的dna顺序相依吸收转化.pdfVIP
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restriction and sequence alterations affect dna uptake sequence-dependent transformation in neisseria meningitidis限制和序列变化影响脑膜炎奈瑟菌的dna顺序相依吸收转化
Restriction and Sequence Alterations Affect DNA Uptake
Sequence-Dependent Transformation in Neisseria
meningitidis
Ole Herman Ambur1,3*, Stephan A. Frye1,2, Mariann Nilsen1,2, Eirik Hovland 1,3, Tone Tønjum1,2,3
1 Centre for Molecular Biology and Neuroscience, Oslo, Norway, 2 Department of Microbiology, Oslo University Hospital (Rikshospitalet), Oslo, Norway, 3 Institute of Basic
Medical Sciences, University of Oslo, Oslo, Norway
Abstract
Transformation is a complex process that involves several interactions from the binding and uptake of naked DNA to
homologous recombination. Some actions affect transformation favourably whereas others act to limit it. Here, meticulous
manipulation of a single type of transforming DNA allowed for quantifying the impact of three different mediators of
meningococcal transformation: NlaIV restriction, homologous recombination and the DNA Uptake Sequence (DUS). In the
wildtype, an inverse relationship between the transformation frequency and the number of NlaIV restriction sites in DNA
was observed when the transforming DNA harboured a heterologous region for selection (ermC) but not when the
transforming DNA was homologous with only a single nucleotide heterology. The influence of homologous sequence in
transforming DNA was further studied using plasmids with a small interruption or larger deletions in the recombinogenic
region and these alterations were found to impair transformation frequency. In contrast, a particularly potent positive driver
of DNA uptake in Neisseria sp. are short DUS in the transforming DNA. However, the molecular mechanism(s) responsible for
DUS specificity remains unknown. Increasing the number of DUS in the transforming DNA was here shown to exert a
positive effect on transformation. Furthermore, an influence of variable placement of DUS relative
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