Recombinant Mesoplasma florum 30S ribosomal protein S5 (rpsE)
Product Specs
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Target Background
Plays a crucial role in translational accuracy, interacting with S4 and S12. Located at the rear of the 30S ribosomal subunit body, it stabilizes the head's conformation relative to the body.
KEGG: mfl:Mfl140
STRING: 265311.Mfl140
Q&A
Basic Research Questions
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What is Mesoplasma florum rpsE and why is it significant for research?
Mesoplasma florum 30S ribosomal protein S5 (rpsE) is a critical component of the small ribosomal subunit that works with S4 and S12 to ensure translational accuracy during protein synthesis . M. florum represents a compelling model organism for systems and synthetic biology due to its near-minimal genome (~800 kb), fast growth rate, and absence of pathogenic potential . The rpsE protein belongs to the universal ribosomal protein uS5 family, making it an important subject for studying fundamental translation mechanisms in one of the simplest free-living organisms.
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How is rpsE positioned within the M. florum genome and transcriptional organization?
The rpsE gene (identifier YP_053380.1, BBF10K_001373) is located within a critical ribosomal protein cluster in the M. florum genome . Transcriptomic analyses reveal that rpsE is co-transcribed with other ribosomal components, including rplO (50S ribosomal protein L15) and secY (preprotein translocase subunit) . This organization reflects the coordinated expression of proteins involved in translation machinery assembly. Directional RNA sequencing and 5'-RACE analyses have identified potential promoter regions and transcription start sites (TSS) controlling rpsE expression, with motif-associated gTSSs showing sharp increases in RNA-seq signal intensity .
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What methods are available for recombinant expression of M. florum rpsE?
Recombinant expression of M. florum rpsE can be achieved using the genetic tools developed specifically for this organism. Three primary transformation methods have been established:
Transformation Method Efficiency (transformants/viable cell) Key Considerations PEG-mediated transformation ~4.1 × 10^-6 Standard protocol, requires more material and hands-on time Electroporation Up to 7.87 × 10^-6 Higher efficiency, requires less material Conjugation from E. coli Up to 8.44 × 10^-7 Novel method, first example of plasmid conjugation from E. coli to Mollicutes For heterologous expression, the E. coli codon-optimized version of rpsE is available (BBF10K_001373) to overcome codon usage bias . Expression systems should incorporate appropriate antibiotic selection markers, with demonstrated functionality for tetracycline, puromycin, and spectinomycin/streptomycin resistance genes in M. florum .
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What is known about the interaction network of rpsE in M. florum?
Protein interaction network analysis reveals that M. florum rpsE engages in high-confidence interactions (score >0.99) with multiple ribosomal proteins :
Interaction Partner Description Functional Relationship rpsF 30S ribosomal protein S6 Binds together with S18 to 16S ribosomal RNA rpsJ 30S ribosomal protein S10 Involved in tRNA binding to ribosomes rplD 50S ribosomal protein L4 Primary rRNA binding protein rpsS 30S ribosomal protein S19 Forms complex with S13 rpsC 30S ribosomal protein S3 Binds mRNA in the 70S ribosome rpsQ 30S ribosomal protein S17 Primary rRNA binding protein rpsZ 30S ribosomal protein S14 Required for 30S particle assembly rpsH 30S ribosomal protein S8 Coordinates assembly of 30S subunit platform These interactions highlight rpsE's role in ribosome structure and function coordination .
