Recombinant Janthinobacterium sp. Ribosome-recycling factor (frr)
Description
Cloning and Expression of Recombinant RRF
Studies on heterologous RRF expression provide a model for Janthinobacterium sp. RRF production:
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Expression Systems: E. coli is commonly used for recombinant RRF production. For example, spinach chloroplast RRF expressed in E. coli exhibited bactericidal activity against RRF-deficient strains .
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Purification: His-tagged RRFs (e.g., from P. aeruginosa) are purified via nickel-affinity chromatography, yielding functional proteins active in ribosome recycling assays .
Key Steps in Recombinant RRF Production:
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Amplification of frr from Janthinobacterium sp. genomic DNA.
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Cloning into expression vectors (e.g., pET15b with N-terminal His-tag).
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Induction with IPTG and purification via immobilized metal affinity chromatography (IMAC).
Functional Analysis and Applications
In Vitro Activity:
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P. aeruginosa RRF successfully dissociated E. coli polysomes into 70S ribosomes in the presence of EF-G, demonstrating cross-species functionality .
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Recombinant spinach RRF disrupted plasmid maintenance in E. coli, highlighting its ribosome-recycling efficiency .
Antibiotic Overproduction Link:
In Streptomyces coelicolor, elevated RRF levels due to frr upregulation enhanced late-phase protein synthesis, directly correlating with actinorhodin overproduction . This suggests recombinant Janthinobacterium sp. RRF could similarly influence secondary metabolite synthesis.
Comparative Genomic and Functional Insights
Table 2: RRF Functional Conservation Across Species
Research Gaps and Future Directions
Product Specs
Target Background
KEGG: mms:mma_2055
STRING: 375286.mma_2055
