Recombinant Soil-borne wheat mosaic virus Replicase large subunit (rep), partial

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Cat. No.
BT2467226
Source
Yeast
Synonyms
rep; ORF1-1bis; Replicase large subunit; EC 2.1.1.-; EC 2.7.7.-; EC 2.7.7.48; EC 3.6.4.13; 209 kDa readthrough protein; RNA-directed RNA polymerase) [Cleaved into: Replicase small subunit; EC 2.1.1.-; EC 2.7.7.-; EC 3.6.4.13; 150 kDa protein; Methyltransferase/RNA helicase)]
Purity
>85% (SDS-PAGE)
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Cat. No.
BT2467226
Source
E.coli
Synonyms
rep; ORF1-1bis; Replicase large subunit; EC 2.1.1.-; EC 2.7.7.-; EC 2.7.7.48; EC 3.6.4.13; 209 kDa readthrough protein; RNA-directed RNA polymerase) [Cleaved into: Replicase small subunit; EC 2.1.1.-; EC 2.7.7.-; EC 3.6.4.13; 150 kDa protein; Methyltransferase/RNA helicase)]
Purity
>85% (SDS-PAGE)
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Cat. No.
BT2467226
Source
E.coli
Synonyms
rep; ORF1-1bis; Replicase large subunit; EC 2.1.1.-; EC 2.7.7.-; EC 2.7.7.48; EC 3.6.4.13; 209 kDa readthrough protein; RNA-directed RNA polymerase) [Cleaved into: Replicase small subunit; EC 2.1.1.-; EC 2.7.7.-; EC 3.6.4.13; 150 kDa protein; Methyltransferase/RNA helicase)]
Purity
>85% (SDS-PAGE)
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Cat. No.
BT2467226
Source
Baculovirus
Synonyms
rep; ORF1-1bis; Replicase large subunit; EC 2.1.1.-; EC 2.7.7.-; EC 2.7.7.48; EC 3.6.4.13; 209 kDa readthrough protein; RNA-directed RNA polymerase) [Cleaved into: Replicase small subunit; EC 2.1.1.-; EC 2.7.7.-; EC 3.6.4.13; 150 kDa protein; Methyltransferase/RNA helicase)]
Purity
>85% (SDS-PAGE)
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Cat. No.
BT2467226
Source
Mammalian cell
Synonyms
rep; ORF1-1bis; Replicase large subunit; EC 2.1.1.-; EC 2.7.7.-; EC 2.7.7.48; EC 3.6.4.13; 209 kDa readthrough protein; RNA-directed RNA polymerase) [Cleaved into: Replicase small subunit; EC 2.1.1.-; EC 2.7.7.-; EC 3.6.4.13; 150 kDa protein; Methyltransferase/RNA helicase)]
Purity
>85% (SDS-PAGE)
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Description

Key Features of the Native Replicase Large Subunit

PropertyDescriptionSource
Genomic LocationEncoded by RNA1 (approx. 7,100 nucleotides)
Molecular Weight~209 kDa (part of RNA1-encoded polyprotein)
Functional RoleCatalyzes viral RNA synthesis; interacts with viral RNA2 and movement proteins
Domain StructureContains conserved motifs for RNA polymerization (e.g., palm, fingers)

Role in Viral Replication

The replicase large subunit is essential for SBWMV’s replication machinery. It forms a heterodimer with the smaller 150 kDa subunit (encoded by RNA1) to synthesize viral RNA from templates . Studies using infectious clones (e.g., agrobacterium-delivered SBWMV cDNA) have demonstrated that mutations in this subunit can disrupt replication fidelity or systemic movement, highlighting its importance in pathogenicity .

Partial Recombinant Constructs in Functional Studies

Partial recombinant constructs are often engineered to:

  1. Map Critical Domains: Truncated versions enable identification of motifs required for RNA binding, catalysis, or interaction with auxiliary proteins (e.g., movement proteins) .

  2. Study RNA Silencing Suppression: SBWMV’s cysteine-rich protein (19K) suppresses host RNA interference, but the replicase may also contribute to this process .

  3. Develop Diagnostic Tools: Recombinant fragments are used in serological assays to detect viral antigens in host tissues .

Infectious Clone Systems

A full-length infectious clone of SBWMV (pSBWMV) was developed using Agrobacterium tumefaciens delivery systems. While the replicase large subunit was not explicitly modified in this study, the clone enables targeted mutagenesis to study its function . For example:

  • Gene Silencing: Insertions into RNA2 (e.g., gene silencing cassettes) were tested for stability and effect on replication .

  • Host-Pathogen Interactions: The clone facilitates studies on how environmental factors (e.g., temperature) influence replicase activity and symptom development .

Synergistic Virus Interactions

In mixed infections with other viruses (e.g., Wheat Streak Mosaic Virus or Brome Mosaic Virus), SBWMV’s replicase may compete for host resources. For instance:

  • Resource Competition: Robust replication of Brome Mosaic Virus suppresses Barley Stripe Mosaic Virus accumulation, suggesting similar dynamics could affect SBWMV replicase activity .

  • RNA Silencing Suppression: The 19K protein of SBWMV counteracts host defenses, potentially enhancing replication efficiency .

Technical Limitations

  1. Stability of Partial Constructs: Truncated replicase subunits may lack essential structural motifs, limiting functional analysis .

  2. Environmental Dependencies: SBWMV replication is temperature-sensitive, complicating in vitro studies .

Emerging Research Priorities

  1. Structural Characterization: Cryo-EM or crystallography studies to resolve the 3D structure of the replicase-subunit complex.

  2. Host-Pathogen Co-Evolution: Investigating how cultivar resistance genes (e.g., Sbwm1) disrupt replicase activity .

Comparative Analysis of SBWMV Replicase with Related Viruses

FeatureSBWMV Replicase Large SubunitBarley Yellow Mosaic Virus (BaYMV)Brome Mosaic Virus (BMV)
Genomic RNABipartite (RNA1/RNA2)Bipartite (RNA1/RNA2)Tripartite (RNA1/RNA2/RNA3)
Replicative StrategySemi-conservative replicationSemi-conservative replicationPositive-sense RNA replication
RNA Silencing Suppression19K CRP (RNA2)VPg (RNA1)1a protein
Temperature SensitivitySystemic movement <20°CN/AN/A

Data synthesized from .

Product Specs

Form
Lyophilized powder
Note: While we prioritize shipping the format currently in stock, please specify your format preference in order notes for customized fulfillment.
Lead Time
Delivery times vary depending on the purchasing method and location. Please contact your local distributor for precise delivery estimates.
Note: Standard shipping includes blue ice packs. Dry ice shipping requires prior arrangement and incurs additional charges.
Notes
Avoid repeated freeze-thaw cycles. Store working aliquots at 4°C for up to one week.
Reconstitution
Centrifuge the vial briefly before opening to collect the contents. Reconstitute the protein in sterile, deionized water to a concentration of 0.1-1.0 mg/mL. We recommend adding 5-50% glycerol (final concentration) and aliquoting for long-term storage at -20°C/-80°C. Our standard glycerol concentration is 50%, which serves as a guideline.
Shelf Life
Shelf life depends on various factors including storage conditions, buffer components, temperature, and the protein's inherent stability. Generally, liquid formulations have a 6-month shelf life at -20°C/-80°C, while lyophilized forms have a 12-month shelf life at -20°C/-80°C.
Storage Condition
Upon receipt, store at -20°C/-80°C. Aliquot for multiple uses to prevent repeated freeze-thaw cycles.
Tag Info
Tag type is determined during manufacturing.
The tag type is determined during production. If a specific tag is required, please inform us, and we will prioritize its inclusion.
Synonyms
rep; ORF1-1bis; Replicase large subunit; EC 2.1.1.-; EC 2.7.7.-; EC 2.7.7.48; EC 3.6.4.13; 209 kDa readthrough protein; RNA-directed RNA polymerase) [Cleaved into: Replicase small subunit; EC 2.1.1.-; EC 2.7.7.-; EC 3.6.4.13; 150 kDa protein; Methyltransferase/RNA helicase)]
Buffer Before Lyophilization
Tris/PBS-based buffer, 6% Trehalose.
Datasheet
Please contact us to get it.
Protein Length
Partial
Purity
>85% (SDS-PAGE)
Species
Soil-borne wheat mosaic virus (strain United States/Nebraska/1981) (SBWMV)
Target Names
rep
Uniprot No.
Q89249

Target Background

Function
This protein is an RNA-dependent RNA polymerase involved in viral RNA replication; it also functions as a methyltransferase in RNA capping and as an RNA helicase. The methyltransferase activity exhibits cytoplasmic capping enzyme function, crucial because all viral RNAs are synthesized in the cytoplasm, unlike host capping enzymes which are nucleus-restricted. The helicase region likely possesses NTPase and RNA unwinding activities (Potential).
Database Links

KEGG: vg:991048

Protein Families
SsRNA positive-strand viruses RNA-directed RNA polymerase family

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