Recombinant Methanocaldococcus jannaschii Uncharacterized protein MJ0110 (MJ0110)
Description
Production and Handling Guidelines
MJ0110 is synthesized via recombinant expression in E. coli, followed by purification and lyophilization. Critical handling parameters include:
Genomic and Functional Context
MJ0110 is encoded by the MJ0110 gene in M. jannaschii, a model organism for studying archaeal metabolism and extremophily. Key genomic insights:
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Genome Status: M. jannaschii was the first archaeon to have its genome sequenced (1996), revealing 1,882 genes, with >30% remaining uncharacterized .
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Pathway Potential: While MJ0110’s role is unknown, M. jannaschii is pivotal in methanogenesis, cofactor biosynthesis, and novel amino acid pathways .
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Uncharacterized Genes: Over a third of M. jannaschii’s genome lacks functional annotation, highlighting MJ0110’s potential as a target for future studies .
Research Applications and Challenges
MJ0110’s recombinant form enables experimental approaches to elucidate its function:
Challenges:
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Absence of high-throughput data (e.g., proteomic interactions) .
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Need for targeted mutagenesis to infer roles in methanogenesis or stress responses .
Future Directions
Ongoing efforts to annotate M. jannaschii’s genome, such as the MjCyc pathway-genome database, aim to resolve uncharacterized genes like MJ0110 . Potential areas of investigation include:
Product Specs
Note: We will prioritize shipping the format we have in stock. However, if you have specific format requirements, please indicate them in your order. We will fulfill your request if possible.
Note: Our proteins are shipped with standard blue ice packs. If you require dry ice shipping, please communicate this to us in advance, as additional fees will apply.
Generally, the shelf life of liquid form is 6 months at -20°C/-80°C. The shelf life of lyophilized form is 12 months at -20°C/-80°C.
Target Background
KEGG: mja:MJ_0110
STRING: 243232.MJ_0110
Q&A
What is the structural and functional context of MJ0110 in Methanocaldococcus jannaschii?
MJ0110, an uncharacterized protein from the thermophilic methanogen Methanocaldococcus jannaschii, is part of a genome known for its unique metabolic pathways and hydrogenase systems . While its exact function remains unclear, its sequence (MVINMDFDITVIGYIAGTLTTFASLPQLIKSLKEKDMSNISLAFVITFTTGLTLWLIYGI LRNDYPIIVFNILSLMFWIPITYLKIRDEMRKS) suggests potential involvement in stress response or metabolic regulation, given the organism’s reliance on hydrogen and carbon dioxide for energy . Recombinant MJ0110 is expressed in E. coli with an N-terminal His-tag, enabling affinity purification, though its thermal stability and native folding require further validation .
How is recombinant MJ0110 produced and purified?
MJ0110 is synthesized via heterologous expression in E. coli, leveraging the organism’s well-characterized genetic machinery . Key steps include:
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Cloning: Insertion of the mj0110 gene into a plasmid vector.
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Induction: IPTG-driven expression under optimized temperature and growth conditions.
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Purification: Ni-NTA affinity chromatography exploiting the His-tag, followed by buffer exchange and lyophilization .
Critical considerations include minimizing denaturation during recombinant expression, as M. jannaschii proteins often require high-temperature renaturation. Post-purification analysis typically involves SDS-PAGE and Western blotting to confirm size and tag integrity .
What challenges exist in studying MJ0110’s functional role?
Three primary hurdles complicate MJ0110 research:
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Thermal Stability: Native M. jannaschii proteins are adapted to high temperatures (e.g., 80–100°C), requiring specialized in vitro assays to mimic physiological conditions .
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Lack of Functional Data: No homologs with known activity exist in public databases, necessitating hypothesis-driven approaches like:
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Experimental Design: Cross-stress exposure studies (e.g., combining heat and oxidative stress) may reveal latent functions, as seen in E. coli biocide-antibiotic interactions .
How should experiments be designed to elucidate MJ0110’s role?
A phased approach leveraging optimal experimental design (OPEX) principles is recommended:
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Hypothesis Generation:
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Phase 1: Broad screening of MJ0110 interactions with metabolic enzymes (e.g., hydrogenases, formate dehydrogenases) using yeast two-hybrid or co-IP.
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Phase 2: High-throughput omics (proteomics, metabolomics) under varied growth conditions (e.g., low H₂, elevated CO₂).
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Model Training:
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Validation:
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Knockout Studies: Generate M. jannaschii Δmj0110 mutants for phenotypic analysis (e.g., growth defects under specific stresses).
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Biochemical Assays: Enzyme activity measurements (e.g., ATP hydrolysis, redox cycling) with purified MJ0110.
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How to resolve contradictions in MJ0110 data analysis?
Conflicting results may arise due to:
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Experimental Artifacts: His-tag interference with native interactions or improper renaturation.
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Data Heterogeneity: Differences in E. coli expression systems vs. native M. jannaschii conditions.
Resolving these requires:
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Meta-Analysis: Cross-referencing MJ0110 studies with other Methanocaldococcus proteins (e.g., hydrogenases) to identify conserved functional motifs .
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Statistical Validation:
What advanced techniques can be used to characterize MJ0110?
How to integrate MJ0110 into broader metabolic models?
For systems biology applications, consider:
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Flux Balance Analysis (FBA): Model MJ0110 as a regulatory node in hydrogen metabolism, using genome-scale reconstructions of M. jannaschii .
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Optimal Experimental Design (OPEX): Prioritize experiments that reduce uncertainty in MJ0110’s role, such as testing its expression under anaerobic vs. microaerobic conditions .
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Cross-Stress Analysis: Investigate MJ0110’s potential role in cross-protection mechanisms (e.g., linking heat shock and oxidative stress responses) .
What are the limitations of current MJ0110 research?
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Structural Gaps: No crystallographic or cryo-EM data exist for MJ0110, limiting mechanistic insights.
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Phylogenetic Bias: Most studies focus on M. jannaschii, neglecting evolutionary conservation across other methanogens.
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Functional Redundancy: Overlapping roles with other uncharacterized proteins complicate knockout phenotype interpretation.
