Intra-species tranduction with Proteus mirabilis high frequency transducing phages
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Abstract:
The properties of three additional Proteus mirabilis hosts for the high frequency transducing (HET) phages 5006MHFTkappa and 5006MHFTak are described. The phages transduce resistance to kanamycin and to ampicillin plus kanamycin, respectively, and were produced by ultraviolet induction of derivatives of P. mirabilis strain PM5006. Strain PM804 could not be shown to adsorb the phages although it yielded a few transductants. All transductants, even those produced at low multiplicities of phage input were lysogenic and segregated markers at high frequency. Ultraviolet induced phage lysates of these transductants transduced PM804 at higher frequencies and PM5006 at lower frequencies than the original phages. Strain PM804 or its derivatives did not adsorb phage from these lysates. Tranmission experiments through PM5006 of phage in the transductant lysates confirmed that PM804 had a host-controlled modification system which modified HFT phage from PM5006. That PM804 also possessed a restriction system was inferred from the greater numbers of transductants obtained with page which bore a compatible modification pattern. Strain N and a restrictionless mutant of it named NHI, adsorbed the HET phages avidly and failed to modify their host range. Transduction frequencies of the phage markers were about 10(-4)/p.f.u. adsorbed to strain NHI and only about tenfold lower to strain N which did not plate the phages. Tranductants also had the features of heterogenotes and those obtained at low multiplicites of infection were non-lysogenic. The latter transductants adsorbed homolgous phage while NHI transductants also plated the HFT lysates. Strain NHI, lysogenized by the parent phage 5006M, did not adsorb the HFT phages. These findings suggest that the HFT phages were most likely defective for lysogenic conversion to homologous phage non-adsorption. The postulated restriction enzymes of PM804 and strain N could not be shown to be thermolabile.
Insights
This study characterizes Proteus mirabilis hosts for high frequency transducing (HFT) phages, revealing host-controlled modification and restriction systems. These systems influence phage transduction and host range, suggesting HFT phages may be defective in lysogenic conversion.
Area of Science:
- Microbiology
- Bacteriology
- Virology
Background:
- Proteus mirabilis is a Gram-negative bacterium.
- High frequency transducing (HFT) phages are specialized bacteriophages capable of efficient gene transfer.
- Understanding phage-host interactions is crucial for molecular biology and genetic engineering.
Purpose of the Study:
- To describe the properties of novel Proteus mirabilis hosts for HFT phages 5006MHFTkappa and 5006MHFTak.
- To investigate the mechanisms of host-controlled modification and restriction in P. mirabilis.
- To elucidate the role of these systems in phage transduction and host range.
Main Methods:
- Bacterial strain characterization and phage adsorption assays.
- Transduction experiments to assess gene transfer efficiency.
- Lysogenic conversion and segregation analysis.
- Ultraviolet induction of phages and analysis of transductant properties.
Main Results:
- Two new P. mirabilis hosts, PM804 and strain N, exhibited distinct phage adsorption, modification, and restriction capabilities.
- PM804 demonstrated a host-controlled modification system and a restriction system influencing HFT phage transduction.
- Strain N and its restrictionless mutant NHI showed avid phage adsorption and limited modification, with varying transduction frequencies.
- Transductants exhibited heterogenote features, with non-lysogenic transductants adsorbing homologous phage.
Conclusions:
- The HFT phages studied are likely defective for lysogenic conversion, leading to homologous phage non-adsorption.
- Host-controlled modification and restriction systems in P. mirabilis play significant roles in regulating phage-host interactions.
- Further research is needed to characterize the specific restriction enzymes involved.