%0 Journal Article %1 claus_identification_1998 %A Claus, H %A Frosch, M %A Vogel, U %D 1998 %J Molecular & General Genetics: MGG %K Analysis, Bacterial Bacterial, Base Chain Chromosome Chromosomes, Cloning, Data, Elements, Genetic Mapping, Molecular Molecular, Mutagenesis, Neisseria Polymerase Reaction, Sequence Sequence, Techniques, Transformation, Transposable meningitidis, {DNA} {DNA}, %N 4 %P 363--371 %T Identification of a hotspot for transformation of Neisseria meningitidis by shuttle mutagenesis using signature-tagged transposons %U http://www.ncbi.nlm.nih.gov/pubmed/9790590 %V 259 %X Shuttle mutagenesis using signature-tagged transposons was employed to generate a library of individually tagged mutants of the Neisseria meningitidis strain B1940, which belongs to serogroup B. The use of tagged transposons allowed us to monitor for enrichment for single mutants during the process of shuttle mutagenesis, by amplification of the tags and subsequent sequence determination. Enrichment of a single clone occurred during the transformation of the meningococci with transposon-containing plasmid DNA. Sequence determination around the site of transposon insertion revealed that the transposon had mutagenized a previously unknown locus, which was designated hrtA (high rate of transformation). hrtA-mediated transformation was independent of TnMax5 and tag sequences, and it most probably involved recombination events. The hrtA locus is restricted to meningococci and gonococci and is present in few apathogenic neisserial species. Chromosomal mapping of hrtA and six further hrt sites revealed a random distribution of highly transforming DNA fragments on the meningococcal chromosome. In conclusion, our data demonstrate that shuttle mutagenesis of naturally competent bacteria using signature-tagged transposons allows the isolation of chromosomal DNA fragments, which exhibit a high transformation efficiency, and which, therefore, are likely to be involved in horizontal gene transfer.

@article{claus_identification_1998, abstract = {Shuttle mutagenesis using signature-tagged transposons was employed to generate a library of individually tagged mutants of the Neisseria meningitidis strain B1940, which belongs to serogroup B. The use of tagged transposons allowed us to monitor for enrichment for single mutants during the process of shuttle mutagenesis, by amplification of the tags and subsequent sequence determination. Enrichment of a single clone occurred during the transformation of the meningococci with transposon-containing plasmid {DNA.} Sequence determination around the site of transposon insertion revealed that the transposon had mutagenized a previously unknown locus, which was designated {hrtA} (high rate of transformation). {hrtA-mediated} transformation was independent of {TnMax5} and tag sequences, and it most probably involved recombination events. The {hrtA} locus is restricted to meningococci and gonococci and is present in few apathogenic neisserial species. Chromosomal mapping of {hrtA} and six further hrt sites revealed a random distribution of highly transforming {DNA} fragments on the meningococcal chromosome. In conclusion, our data demonstrate that shuttle mutagenesis of naturally competent bacteria using signature-tagged transposons allows the isolation of chromosomal {DNA} fragments, which exhibit a high transformation efficiency, and which, therefore, are likely to be involved in horizontal gene transfer.}, added-at = {2011-06-24T11:21:47.000+0200}, author = {Claus, H and Frosch, M and Vogel, U}, biburl = {https://www.bibsonomy.org/bibtex/2b0447bb1f389ede24a34d5ef327128f6/ag_vogel}, interhash = {b016ddc445cf7aa4c346ecc2f5f05287}, intrahash = {b0447bb1f389ede24a34d5ef327128f6}, issn = {0026-8925}, journal = {Molecular \& General Genetics: {MGG}}, keywords = {Analysis, Bacterial Bacterial, Base Chain Chromosome Chromosomes, Cloning, Data, Elements, Genetic Mapping, Molecular Molecular, Mutagenesis, Neisseria Polymerase Reaction, Sequence Sequence, Techniques, Transformation, Transposable meningitidis, {DNA} {DNA},}, month = sep, note = {{PMID:} 9790590}, number = 4, pages = {363--371}, timestamp = {2011-06-24T11:21:50.000+0200}, title = {Identification of a hotspot for transformation of Neisseria meningitidis by shuttle mutagenesis using signature-tagged transposons}, url = {http://www.ncbi.nlm.nih.gov/pubmed/9790590}, volume = 259, year = 1998 }