Recombinant Salmonella gallinarum Ribosome-recycling factor (frr)

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Cat. No.
BT45560
Source
Yeast
Synonyms
frr; SG0223; Ribosome-recycling factor; RRF; Ribosome-releasing factor
Purity
>85% (SDS-PAGE)
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Cat. No.
BT45560
Source
E.coli
Synonyms
frr; SG0223; Ribosome-recycling factor; RRF; Ribosome-releasing factor
Purity
>85% (SDS-PAGE)
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Cat. No.
BT45560
Source
E.coli
Synonyms
frr; SG0223; Ribosome-recycling factor; RRF; Ribosome-releasing factor
Purity
>85% (SDS-PAGE)
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Cat. No.
BT45560
Source
Baculovirus
Synonyms
frr; SG0223; Ribosome-recycling factor; RRF; Ribosome-releasing factor
Purity
>85% (SDS-PAGE)
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Cat. No.
BT45560
Source
Mammalian cell
Synonyms
frr; SG0223; Ribosome-recycling factor; RRF; Ribosome-releasing factor
Purity
>85% (SDS-PAGE)
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Description

Biological Role of RRF in Bacteria

RRF collaborates with elongation factor G (EF-G) to split ribosomes into subunits after translation termination, enabling their reuse. Key features include:

  • Structural domains:

    • Domain A: N-terminal helix critical for binding to the 50S ribosomal subunit.

    • Domain B: Central region with β-strand/coil structures essential for structural stability .

    • Domain C: C-terminal helix containing active sites (e.g., Arg132 in E. coli) .

  • Functional impact: Depletion of RRF in E. coli leads to ribosome "traffic jams" at stop codons and aberrant 3′-UTR ribosome accumulation but does not enhance translational coupling .

2.1. Gene Deletion and Complementation Strategies

  • CRISPR/Cas9 and λ-Red recombination: Used in S. gallinarum to delete virulence genes (e.g., SpvB , purB ). Similar methods could target frr.

    • Example protocol for gene deletion:

      1. Transform S. gallinarum with temperature-sensitive plasmid pKD46 (expressing recombinases).

      2. Introduce homologous recombination cassette (e.g., cat gene flanked by frr homology arms).

      3. Remove antibiotic markers via FLP/FRT recombination .

2.2. Plasmid-Based Expression Systems

  • Antigen delivery vectors: S. gallinarum strains like SG100 (Δasd) are engineered to express heterologous antigens (e.g., APEC type I fimbriae ) using plasmids such as pYA3342.

    • Hypothetical RRF expression: A similar approach could clone frr into a shuttle vector (e.g., pBR322) under a regulated promoter for recombinant RRF production.

Comparative Analysis of RRF Across Species

FeatureE. coli RRF Hypothetical S. gallinarum RRF
Gene length198 aa (UniProt P0AGD7)~131 aa (inferred from S. enterica)
Critical residuesArg132 (domain C)Likely conserved
Null mutationsDisrupt ribosome recyclingUntested in S. gallinarum
ThermosensitivityDomain A/C mutations cause ts phenotypes Uncharacterized

Research Gaps and Future Directions

  • Functional studies: No direct evidence exists for frr’s role in S. gallinarum pathogenesis or vaccine efficacy.

  • Expression challenges: Ribosome recycling is essential for viability, necessitating conditional knockdown systems (e.g., arabinose-inducible promoters) to study frr deletion.

  • Applications: Engineered S. gallinarum strains expressing RRF variants could elucidate its role in bacterial fitness or serve as attenuated vaccine platforms.

5.1. Cloning and Purification (Hypothetical Workflow)

  1. Amplify *frr*: Design primers with BamHI/XhoI sites using S. gallinarum genomic DNA.

  2. Ligate into pET-28a: Express His-tagged RRF in E. coli BL21(DE3).

  3. Purify via Ni-NTA:

    • Buffer: 50 mM Tris-HCl, 300 mM NaCl, 250 mM imidazole (pH 8.0).

    • Yield: ~85% purity (similar to YRAN recombinant protein protocols ).

Product Specs

Form
Lyophilized powder. We will ship the in-stock format unless you specify a preference when ordering.
Lead Time
Delivery times vary by purchase method and location. Consult local distributors for specific delivery times. Proteins are shipped with blue ice packs by default. Contact us in advance for dry ice shipping (extra fees apply).
Notes
Avoid repeated freeze-thaw cycles. Store working aliquots at 4°C for up to one week.
Reconstitution
Briefly centrifuge the vial before opening. Reconstitute in sterile deionized water to 0.1-1.0 mg/mL. Add 5-50% glycerol (final concentration) and aliquot for long-term storage at -20°C/-80°C. Our default final glycerol concentration is 50%.
Shelf Life
Shelf life depends on storage conditions, buffer, temperature, and protein stability. Liquid form: 6 months at -20°C/-80°C. Lyophilized form: 12 months at -20°C/-80°C.
Storage Condition
Store at -20°C/-80°C upon arrival. Aliquot for multiple uses. Avoid repeated freeze-thaw cycles.
Tag Info
Tag type is determined during manufacturing. If you require a specific tag, please inform us and we will prioritize its development.
Synonyms
frr; SG0223; Ribosome-recycling factor; RRF; Ribosome-releasing factor
Buffer Before Lyophilization
Tris/PBS-based buffer, 6% Trehalose.
Datasheet
Please contact us to get it.
Expression Region
1-185
Protein Length
full length protein
Purity
>85% (SDS-PAGE)
Species
Salmonella gallinarum (strain 287/91 / NCTC 13346)
Target Names
frr
Target Protein Sequence
MISDIRKDAE VRMEKCVEAF KTQISKVRTG RASPSLLDGI VVEYYGTPTP LRQLASVTVE DSRTLKINVF DRSMGPAVEK AIMASDLGLN PSSAGTDIRV PLPPLTEERR KDLTKIVRGE AEQARVAVRN VRRDANDKVK ALLKDKAISE DDDRRSQEEV QKMTDAAIKK VDAALADKEA ELMQF
Uniprot No.
B5RHF7

Target Background

Function
Releases ribosomes from messenger RNA at the end of protein biosynthesis. May enhance translation efficiency by recycling ribosomes.
Database Links

KEGG: seg:SG0223

Protein Families
RRF family
Subcellular Location
Cytoplasm.

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