Recombinant Bacillus subtilis Uncharacterized protein ywoF (ywoF)

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Cat. No.

BT32649

Source

Yeast

Synonyms

ywoF; BSU36460; Uncharacterized protein YwoF

Purity

>85% (SDS-PAGE)

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Cat. No.

BT32649

Source

E.coli

Synonyms

ywoF; BSU36460; Uncharacterized protein YwoF

Purity

>85% (SDS-PAGE)

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Cat. No.

BT32649

Source

E.coli

Synonyms

ywoF; BSU36460; Uncharacterized protein YwoF

Purity

>85% (SDS-PAGE)

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Cat. No.

BT32649

Source

Baculovirus

Synonyms

ywoF; BSU36460; Uncharacterized protein YwoF

Purity

>85% (SDS-PAGE)

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Cat. No.

BT32649

Source

Mammalian cell

Synonyms

ywoF; BSU36460; Uncharacterized protein YwoF

Purity

>85% (SDS-PAGE)

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Description

Overview of Uncharacterized Proteins in Bacillus subtilis

Uncharacterized proteins in B. subtilis are typically annotated as hypothetical or putative gene products with undetermined functions. These proteins often lack experimental validation but may share structural or sequence similarities with known proteins. For example:

YwsC: A key enzyme in γ-polyglutamate (PGA) biosynthesis, essential for polymer synthesis and secretion .
YwdE: A membrane-associated protein with an uncharacterized role, expressed recombinantly for structural studies .
YwtA: Required for PGA transport across the cell membrane .

Recombinant Protein Production Systems in B. subtilis

While ywoF is not described, the provided studies highlight methodologies for recombinant protein expression in B. subtilis, which could apply to ywoF characterization:

Table 1: Expression Systems for Recombinant Proteins in B. subtilis

Promoter Type	Inducer	Host Strain	Example Protein Expressed	Yield/Activity
Constitutive (P43)	None	WB800N	Trehalose synthase	23,080.6 U/L
Inducible (P grac)	IPTG	SCK6/WB600	rhBMP2	200 ng/mL
Autoinducible	Self-inducing	168 (ATCC 33,712)	Microbial transglutaminase	63.0 ± 0.6 mg/L

Key techniques include:

Signal peptide optimization for secretion (e.g., SPsacB, SPamy) .
Dual-promoter systems to enhance expression levels .
CRISPR/Cas9-mediated gene editing for strain optimization .

Genomic Context of ywoF

Though ywoF is not discussed in the provided sources, analogous uncharacterized genes (e.g., ywsC, ywdE) often reside in operons linked to stress responses or metabolite synthesis. For example:

The ywsC-ywtABC operon is critical for PGA biosynthesis .
ywdE is part of a genomic cluster with unknown regulatory elements .

Research Recommendations for ywoF

To characterize ywoF, the following approaches are suggested:

Heterologous Expression: Clone ywoF into plasmids (e.g., pHT43 or pMA5) under inducible promoters (e.g., Pgrac) .
Proteomic Analysis: Use Western blotting and mass spectrometry to confirm expression and post-translational modifications .
Functional Screens: Test knockout strains for phenotypic changes under stress (e.g., cell wall antibiotics, heat shock) .
Structural Studies: Resolve the protein’s 3D structure via X-ray crystallography or cryo-EM .

Challenges and Limitations

Annotation Gaps: ywoF may be misannotated or species/strain-specific.
Functional Redundancy: Overlap with other ECF σ factor regulons (e.g., σM, σW) could obscure its role .
Secretion Efficiency: Without a validated signal peptide, extracellular yield may be low .

Product Specs

Form

Lyophilized powder. We will ship the format we have in stock. If you have special format requirements, please note them when ordering.

Lead Time

Delivery time varies by purchase method and location. Consult local distributors for specific times. Proteins are shipped with blue ice packs by default. Request dry ice in advance for an extra fee.

Notes

Avoid repeated freezing and thawing. Working aliquots can be stored at 4°C for up to one week.

Reconstitution

Briefly centrifuge the vial before opening. Reconstitute protein 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 ingredients, 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 receipt. 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

ywoF; BSU36460; Uncharacterized protein YwoF

Buffer Before Lyophilization

Tris/PBS-based buffer, 6% Trehalose.

Datasheet

Please contact us to get it.

Expression Region

28-468

Protein Length

Full Length of Mature Protein

Purity

>85% (SDS-PAGE)

Species

Bacillus subtilis (strain 168)

Target Names

ywoF

Target Protein Sequence

NEE GSGDYLYVSP NGSDQNEGTK EKPFRTLAHA SEKAAAGTTV MIREGTYHET LDVKHSGTDG KPITFRNYEN ENVVISGESV ANAEYETPLI RIHDKHDIAI SGLTIQDLSV SSEEATAIGI YVSGSSSHIA IKDNHIRGIK TTADEGNAHG IAVYGTGSMK DIRIEDNTVE KLTLGASEAV VLNGNIDGFT VAGNVVRNNN NIGIDLIGYE GTADKNDYVR NGVVENNTVY QNSTYGNPAY GDEYSAGGIY VDGGHDIEIK NNTVYDNDIG IEATSEHKGK YANAIQITDN KVYNNAYTGI SIGGYDKKRG GTSNSLIARN IMYRNDTKGL YGGQLLLQYD TKNNTIEKNI LTAGDSRLFI GNDFTENEGN TVNHNVYHKE ADQDGIWMWK KKEYDSFSSY RKATKNDQQS IYADPMFRDE ASYDFSLDPD SPARPVIE

Uniprot No.

P94576

Target Background

Database Links

KEGG: bsu:BSU36460

STRING: 224308.Bsubs1_010100019706

Subcellular Location

Secreted.

Q&A

Since the provided search results do not contain specific information about the uncharacterized protein YwoF in Bacillus subtilis, the following FAQs are constructed based on general methodologies for studying uncharacterized proteins in recombinant B. subtilis systems, informed by broader research practices in microbial genetics and protein characterization. Below is a framework that adheres to academic research standards and the requirements outlined:

How to design experiments for initial characterization of uncharacterized YwoF in recombinant B. subtilis?

Methodological Answer:

Bioinformatic Analysis: Perform sequence alignment (BLAST, Pfam) to identify conserved domains and potential homologs. Use tools like AlphaFold2 to predict tertiary structure.
Expression Optimization: Use inducible promoters (e.g., P $_{grac}$ or xylose-inducible systems) to express YwoF in B. subtilis . Monitor expression via SDS-PAGE and Western blot.
Knockout Strains: Generate ΔywoF mutants via homologous recombination (e.g., using pMAD or CRISPR-Cas9 systems) and compare growth phenotypes under stress (e.g., DNA damage, nutrient limitation) .

What experimental strategies confirm the subcellular localization of YwoF?

Methodological Answer:

Fractionation: Separate cytoplasmic, membrane, and extracellular fractions via ultracentrifugation. Validate purity using marker proteins (e.g., GroEL for cytoplasm, SecA for membrane) .
Fluorescent Tagging: Fuse YwoF with GFP under a constitutive promoter (e.g., P $_{veg}$ ) and visualize localization via fluorescence microscopy .
Secretome Analysis: Use LC-MS/MS to detect YwoF in extracellular proteome samples .

How to resolve contradictions in functional data for YwoF across studies?

Methodological Answer:

Condition-Specific Assays: Test YwoF activity under varying conditions (e.g., pH, temperature, oxidative stress) to identify context-dependent roles.
Genetic Interaction Mapping: Perform synthetic lethality screens with ΔywoF and mutants of related pathways (e.g., DNA repair genes recA, ruvA) .
Multi-Omics Integration: Correlate transcriptomic (RNA-seq) and proteomic (TMT labeling) data from ΔywoF strains to identify dysregulated pathways .

What advanced techniques determine YwoF's role in chromosomal segregation or DNA repair?

Methodological Answer:

Single-Molecule Imaging: Use fluorescent lacO arrays to visualize chromosome segregation defects in ΔywoF strains .
Pull-Down Assays: Identify YwoF interaction partners (e.g., RecU, RuvAB) using His-tagged YwoF and Ni-NTA affinity chromatography .
Complementation Studies: Express YwoF orthologs from related species (e.g., B. licheniformis) in ΔywoF strains to assess functional conservation .

Table 1: Genetic Tools for YwoF Expression in B. subtilis

Tool Type	Example Systems	Application
Inducible Promoters	P $_{grac}$ , P $_{xyl}$	Tight regulation of YwoF expression
Secretion Vectors	pHT43 (SP $_{lipA}$ signal)	Extracellular YwoF production
Knockout Systems	CRISPR-Cas9, pMAD	Generation of ΔywoF mutants

Table 2: Analytical Techniques for YwoF Characterization

Technique	Key Parameters	Use Case
2D Gel Electrophoresis	pI, molecular weight	Detect post-translational modifications
Surface Plasmon Resonance (SPR)	Binding kinetics (k $_{on}$ /k $_{off}$ )	Measure interactions with DNA or proteins
Cryo-EM	Resolution < 3.5 Å	Resolve YwoF structure in complex with partners

Key Methodological Challenges

Low Abundance: Use tandem affinity purification (TAP) with dual tags (e.g., Strep-II/His $_6$ ) to enhance YwoF recovery .
Functional Redundancy: Perform high-throughput suppressor screens to identify compensatory pathways in ΔywoF strains .
Data Reproducibility: Standardize growth conditions (e.g., Schaeffer’s sporulation medium vs. LB) to minimize variability .

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Recombinant Bacillus subtilis Uncharacterized protein ywoF (ywoF)

Description

Overview of Uncharacterized Proteins in Bacillus subtilis

Recombinant Protein Production Systems in B. subtilis

Table 1: Expression Systems for Recombinant Proteins in B. subtilis

Genomic Context of ywoF

Research Recommendations for ywoF

Challenges and Limitations

Product Specs

Target Background

Q&A

How to design experiments for initial characterization of uncharacterized YwoF in recombinant B. subtilis?

What experimental strategies confirm the subcellular localization of YwoF?

How to resolve contradictions in functional data for YwoF across studies?

What advanced techniques determine YwoF's role in chromosomal segregation or DNA repair?

Table 1: Genetic Tools for YwoF Expression in B. subtilis

Table 2: Analytical Techniques for YwoF Characterization

Key Methodological Challenges

Quick Inquiry

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