ArticleApplied and environmental microbiology2005
Generation and molecular characterization of new temperature-sensitive plasmids intended for genetic engineering of Pasteurellaceae.
Article in Applied and environmental microbiology, 2005. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed, 29 citations in OpenAlex.
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- Pasteurella multocida filamentous hemagglutinin B1 (fhaB1) gene is not involved with avian fowl cholera pathogenesis in turkey poults.BMC veterinary research · 2025Article
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- A marker-free genetic manipulation method for Glaesserella parasuis strains developed by alternately culturing transformants at 37°C and 30°C.BMC biotechnology · 2024Article
- Efficient markerless genetic manipulation ofApplied and environmental microbiology · 2024Article
- Generation of markerless and multiple-gene knockout in Glaesserella parasuis based on natural transformation and Flp recombinase.Applied microbiology and biotechnology · 2022Article
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- The prokaryotic Argonaute proteins enhance homology sequence-directed recombination in bacteria.Nucleic acids research · 2019Article
- Article
- Computational design and characterization of a temperature-sensitive plasmid replicon for gram positive thermophiles.Journal of biological engineering · 2012Article
- Construction of non-polar mutants in Haemophilus influenzae using FLP recombinase technology.BMC molecular biology · 2008Article
- Construction of in-frame aroA deletion mutants of Mannheimia haemolytica, Pasteurella multocida, and Haemophilus somnus by using a new temperature-sensitive plasmid.Applied and environmental microbiology · 2005Article
Corrections and comments
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Authors and funding
2 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Temperature-sensitive (TS) plasmids were generated through chemical mutagenesis of a derivative of the streptomycin resistance parent plasmid pD70, isolated from Mannheimia hemolytica serotype 1. Three TS plasmids which failed to replicate at or above 42 degrees C in M. hemolytica but which were fully functional below 31 degrees C were selected for further analysis. Two of the TS plasmids were shown by sequencing to possess unique single-base-pair mutations. The third TS plasmid contained a unique base pair substitution and a second mutation that had been previously identified. These mutations were clustered within a 200-bp region of the presumed plasmid origin of replication. Site-directed single-nucleotide substitutions were introduced into the wild-type pD70 origin of replication to confirm that mutations identified by sequencing had conferred thermoregulated replication. Deletion analysis on the wild-type pD70 plasmid replicon revealed that approximately 720 bp are necessary for plasmid maintenance. Replication of the TS plasmids was thermoregulated in Pasteurella multocida and Haemophilus somnus as well. To consistently transform H. somnus with TS plasmid, in vitro DNA methylation with commercially available HhaI methyltransferase was necessary to protect against the organism's restriction enzyme HsoI (recognition sequence 5'-GCGC-3') characterized herein.
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