ArticleProceedings of the National Academy of Sciences of the United States of America2009
Transformation and isolation of allelic exchange mutants of Chlamydia psittaci using recombinant DNA introduced by electroporation.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 59 papers.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
59 citing papers in PubMed, 91 citations in OpenAlex.
- Article
- Zoonotic and other veterinary chlamydiae - an update, the role of the plasmid and plasmid-mediated transformation.Pathogens and disease · 2024Review
- Advances in genetic manipulation ofFrontiers in immunology · 2023Review
- Recent advances in genetic systems in obligate intracellular human-pathogenic bacteria.Frontiers in cellular and infection microbiology · 2023Review
- Targeted mutagenesis in Anaplasma marginale to define virulence and vaccine development against bovine anaplasmosis.PLoS pathogens · 2022Article
- The Impact of Lateral Gene Transfer inFrontiers in cellular and infection microbiology · 2022Review
- Analysis of Orientia tsutsugamushi promoter activity.Pathogens and disease · 2021Article
- The growing repertoire of genetic tools for dissecting chlamydial pathogenesis.Pathogens and disease · 2021Article
- A broad-spectrum cloning vector that exists as both an integrated element and a free plasmid in Chlamydia trachomatis.PloS one · 2021Article
- An inducible transposon mutagenesis approach for the intracellular human pathogenWellcome open research · 2021Article
- Development of a Plasmid Shuttle Vector System for Genetic Manipulation of Chlamydia psittaci.mSphere · 2020Article
- Gene transfer to plants by electroporation: methods and applications.Molecular biology reports · 2020Review
- Diversity in Chlamydial plasmids.PloS one · 2020Article
- Article
- Chlamydial Infection From Outside to Inside.Frontiers in microbiology · 2019Review
- The Expanding Molecular Genetics Tool Kit in Chlamydia.Journal of bacteriology · 2018Article
- Fluorescence-Reported Allelic Exchange Mutagenesis Reveals a Role for Chlamydia trachomatis TmeA in Invasion That Is Independent of Host AHNAK.Infection and immunity · 2017Article
- A genetic system for targeted mutations to disrupt and restore genes in the obligate bacterium, Ehrlichia chaffeensis.Scientific reports · 2017Article
- Engineering of obligate intracellular bacteria: progress, challenges and paradigms.Nature reviews. Microbiology · 2017Review
- GFPuv-Expressing Recombinant Rickettsia typhi: a Useful Tool for the Study of Pathogenesis and CD8Infection and immunity · 2017Article
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
Abstract
To facilitate genetic investigations in the obligate intracellular pathogens Chlamydia, the ability to construct variants by homologous recombination was investigated in C. psittaci 6BC. The single rRNA operon was targeted with a synthetic 16S rRNA allele, harboring three nucleotide substitutions over 398 bp, which imparts resistance to kasugamycin (Ksm) and spectinomycin (Spc) and causes loss of one HpaI restriction site. A fourth, silent mutation was introduced 654 bp downstream in the beginning of the 23S rRNA gene. C. psittaci 6BC infectious particles were electroporated with various concentrations of circular or linearized plasmids containing different lengths of the rRNA region homologous to the chromosomal copy except for the four nucleotide substitutions. Ksm and Spc were added 18 h after inoculation onto confluent cell monolayers in the plaque assay. Resistant plaques were picked and expanded with selection 10 days later before collecting DNA for analysis by PCR, restriction mapping, sequencing, or Southern. Spontaneous resistance to Ksm and Spc was never observed in mock electroporated bacteria (frequency <6.2 x 10(-9)). Conversely, double resistance and replacement of the 16S rRNA gene were observed when C. psittaci was electroporated with the recombination substrates. Highest efficiency was obtained with 10 microg of circular vector prepared in a DNA methylase-deficient Escherichia coli (1.9 +/- 1.1 x 10(-6), n = 7). Coinheritance of the silent 23S rRNA mutation was seen in 46 of 67 recombinants analyzed, illustrating DNA exchange of up to 1,052 bp in length. These findings provide the first step toward genetic manipulation of Chlamydia.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.