ArticlePloS one2011
Development of shuttle vectors for transformation of diverse Rickettsia species.
Article in PloS one, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 52 papers, 1 of them a synthesis that pooled it.
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Who cites it
52 citing papers in PubMed, 1 synthesis or guideline pooled it.
- Genomic evolution and adaptation of arthropod-associated Rickettsia.Scientific reports · 2022Pooled it
- Transformation and allelic exchange inmBio · 2026Article
- Divergent Rickettsia species exhibit distinct mechanisms of actin-based motility.The Journal of cell biology · 2026Article
- Transformation and allelic exchange inbioRxiv : the preprint server for biology · 2026Article
- Article
- Development of a Recombinase-Mediated Cassette Exchange System for Gene Knockout and Expression of Non-Native Gene Sequences inVaccines · 2025Article
- Development of inducible promoter and CRISPRi plasmids functional inJournal of bacteriology · 2024Article
- An expanded genetic toolkit for inducible expression and targeted gene silencing inJournal of bacteriology · 2024Article
- An expanded genetic toolkit for inducible expression and targeted gene silencing inbioRxiv : the preprint server for biology · 2024Article
- PathogenicJournal of bacteriology · 2024Review
- The role of autophagy in tick-endosymbiont interactions: insights fromMicrobiology spectrum · 2024Article
- Metabolism and physiology of pathogenic bacterial obligate intracellular parasites.Frontiers in cellular and infection microbiology · 2024Review
- Quantitative analysis of morphogenesis and growth dynamics in an obligate intracellular bacterium.Molecular biology of the cell · 2023Article
- Recent advances in genetic systems in obligate intracellular human-pathogenic bacteria.Frontiers in cellular and infection microbiology · 2023Review
- Article
- Bringing genetics to heretofore intractable obligate intracellular bacterial pathogens: Chlamydia and beyond.PLoS pathogens · 2022Article
- Examination of Rickettsial Host Range for Shuttle Vectors Based onApplied and environmental microbiology · 2022Article
- Article
- Cells within cells: Rickettsiales and the obligate intracellular bacterial lifestyle.Nature reviews. Microbiology · 2021Review
- The enigmatic biology of rickettsiae: recent advances, open questions and outlook.Pathogens and disease · 2021Review
Corrections and comments
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Authors and funding
8 authors.
Funding
Abstract
Plasmids have been identified in most species of Rickettsia examined, with some species maintaining multiple different plasmids. Three distinct plasmids were demonstrated in Rickettsia amblyommii AaR/SC by Southern analysis using plasmid specific probes. Copy numbers of pRAM18, pRAM23 and pRAM32 per chromosome in AaR/SC were estimated by real-time PCR to be 2.0, 1.9 and 1.3 respectively. Cloning and sequencing of R. amblyommii AaR/SC plasmids provided an opportunity to develop shuttle vectors for transformation of rickettsiae. A selection cassette encoding rifampin resistance and a fluorescent marker was inserted into pRAM18 yielding a 27.6 kbp recombinant plasmid, pRAM18/Rif/GFPuv. Electroporation of Rickettsia parkeri and Rickettsia bellii with pRAM18/Rif/GFPuv yielded GFPuv-expressing rickettsiae within 2 weeks. Smaller vectors, pRAM18dRG, pRAM18dRGA and pRAM32dRGA each bearing the same selection cassette, were made by moving the parA and dnaA-like genes from pRAM18 or pRAM32 into a vector backbone. R. bellii maintained the highest numbers of pRAM18dRGA (13.3 - 28.1 copies), and R. parkeri, Rickettsia monacensis and Rickettsia montanensis contained 9.9, 5.5 and 7.5 copies respectively. The same species transformed with pRAM32dRGA maintained 2.6, 2.5, 3.2 and 3.6 copies. pRM, the plasmid native to R. monacensis, was still present in shuttle vector transformed R. monacensis at a level similar to that found in wild type R. monacensis after 15 subcultures. Stable transformation of diverse rickettsiae was achieved with a shuttle vector system based on R. amblyommii plasmids pRAM18 and pRAM32, providing a new research tool that will greatly facilitate genetic and biological studies of rickettsiae.
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