Recombinant Bdellovibrio bacteriovorus Elongation factor G 1 (fusA1), partial
Product Specs
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Target Background
KEGG: bba:Bd2979
STRING: 264462.Bd2979
Q&A
FAQs for Researchers Investigating Recombinant Bdellovibrio bacteriovorus Elongation Factor G 1 (fusA1), Partial
How do contradictory data on EF-G1’s role in cell elongation inform experimental design?
Conflicting models of B. bacteriovorus elongation (unipolar vs. dispersed growth ) require:
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High-resolution time-lapse microscopy: Track EF-G1 localization during predatory growth using fluorescent tags (e.g., mTFP fusions ).
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Comparative mutagenesis: Test domain-specific fusA1 mutations (e.g., domain IV Asn592Ile ) for impacts on filamentation kinetics and prey lysis timing.
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Table 1: Key discrepancies and resolution approaches
What controls are critical when assessing fusA1 mutagenesis effects on predatory behavior?
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Prey lysis validation: Confirm bdelloplast integrity via phase-contrast microscopy and nucleic acid release assays .
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Fitness cost analysis: Compare predation rates of mutants vs. wild-type using plaque-forming unit (PFU) assays on GFP-labeled Salmonella prey .
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RT-qPCR normalization: Use rpsG (cotranscribed with fusA1 ) as a reference gene during expression profiling.
How can researchers reconcile EF-G1’s dual roles in translation and cell morphogenesis?
Advanced approaches include:
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Ribosome profiling: Compare translatome changes in fusA1 mutants during intra-bdelloplast growth .
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Cryo-ET imaging: Visualize EF-G1-ribosome interactions in elongating B. bacteriovorus cells .
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ATPase activity assays: Couple GTP hydrolysis rates with predator cell elongation measurements .
What bioinformatic tools are recommended for predicting fusA1 mutation impacts?
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Molecular dynamics simulations: Model domain II/III mutations (e.g., Arg371Cys) to predict structural destabilization .
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Phylogenetic footprinting: Identify conserved residues across Bdellovibrio strains using OrthoFinder or PHYLIP.
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DCA (Direct Coupling Analysis): Detect co-evolving residues in EF-G1 to guide combinatorial mutagenesis .
How do methodological limitations affect fusA1 functional studies?
Key pitfalls and mitigations:
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Axenic growth artifacts: Use prey-dependent predation assays (not HI strains) for phenotypic validation .
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Tag interference: Validate fluorescently tagged EF-G1 via complementation of ΔfusA1 lethality .
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Heterogeneous predation: Standardize prey:predator ratios (10:1) and monitor via real-time prey bioluminescence .
