Recombinant Snakehead rhabdovirus Glycoprotein G (G)
Description
Key Features of Snakehead Rhabdovirus Glycoprotein G (G)
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Viral Glycoprotein: Glycoprotein G is a protein expressed by the snakehead rhabdovirus .
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Role in Autophagy: Rhabdovirus-encoded glycoprotein can induce autophagy in the host by interacting with SnRK1 and enhancing its kinase activity on ATG6 . Glycoproteins can be recognized by ATG6, which acts as a bridge linking glycoproteins to the autophagosome protein ATG8 for degradation .
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Temperature Tolerance: SHRV replicates at high temperatures, which makes it suitable for delivering foreign proteins to fish in warmer waters .
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Recombinant Production: Recombinant SHRV can be generated using reverse genetics to express foreign proteins in fish cells .
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Vaccine Development: Recombinant SHRVs have potential for developing combined vaccines by expressing multiple foreign antigens .
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Protein characteristics: Recombinant full-length Snakehead rhabdovirus Glycoprotein G(G) Protein (Q9QJT6) (26-512aa), fused to N-terminal His tag, can be expressed in E. coli .
Recombinant SHRV Generation
To utilize SHRV as a vehicle for delivering foreign proteins, researchers have developed recombinant SHRVs. For example, a recombinant SHRV containing an enhanced-GFP (eGFP) gene between the nucleoprotein (N) and phosphoprotein (P) genes (rSHRV-A-eGFP) has been created . Another recombinant SHRV expressing two heterologous genes was made by inserting an eGFP gene between N and P genes, and an mCherry gene between P and M genes (rSHRV-AeGFP-BmCherry) .
Applications in Vaccine Development
Recombinant SHRVs can express multiple foreign antigens, making them suitable for combined vaccines .
Impact of Glycoprotein G on Viral Pathogenicity
The glycoprotein G of rhabdoviruses plays a crucial role in viral infection and host immune responses . Research indicates that the level of G incorporation into virions can influence viral pathogenicity .
NV Gene and SHRV Replication
Studies have shown that the nonvirion (NV) gene of SHRV is not essential for viral replication in cultured fish cells . Recombinant viruses with a truncated NV protein can be produced at concentrations similar to wild-type viruses .
Replacement of SHRV Glycoprotein
The SHRV glycoprotein can be replaced by glycoproteins from other viruses, such as the infectious hematopoietic necrosis virus (IHNV), or by hybrid proteins composed of SHRV and IHNV sequences .
Recombinant Glycoprotein G(G) Protein Information
Recombinant Full Length Snakehead rhabdovirus Glycoprotein G(G) Protein, His-Tagged
| Category | Information |
|---|---|
| Cat.No. | RFL6002SF |
| Product Overview | Recombinant Full Length Snakehead rhabdovirus Glycoprotein G(G) Protein (Q9QJT6) (26-512aa), fused to N-terminal His tag, was expressed in E. coli. |
| Species | Snakehead rhabdovirus (SHRV) |
| Source | E. coli |
| Tag | His |
| Protein Length | Full Length of Mature Protein (26-512) |
| Form | Lyophilized powder |
| AA Sequence | QVTHKPRPDSIVEYSEEWENPIYTTPSHCFEDTFAPVKPEKLRCPHIFDDQNLGLTASKA KILHMDLKPEDTHFEAKGRLLHKVTYQVLCSTGFFGGRTVTRKVLETPMGDNEAQAYKAV DREFPYFPEPLCFWLRDNVAAERVFHFSTPKTVTVDLYSRKYISPDFVGGQCAKSPCPTH WPNVYWVGETQSPECPSIDTEGGHIFTKKDTHRITKAVVHGHHPWGLTKACQIQFCNEQW IRTDLGDLIRIEPNDGTSSLTLPKCQDNVVQMRGNLDDFSYLNHAIVNMAQRSECLEAHS SIVAQQKVSPYLLSKFRPPHPGLGKAHYLQNNTIMRGDCIYEGVAEISENRTTYRNLKGE WKKWSLSRGGEGYDGMTVGTKIVIPDLEKYQSIYDNGMFIPKLLGEVPHPSIVITYNQTD SIETGIFTDGKLLNMGVNWTLWPSLSGISLFTVASLILIWYCCCRVTPQALNYSIPMHTI TSRGVEI |
| Purity | Greater than 90% as determined by SDS-PAGE. |
| Storage | Store at -20°C/-80°C upon receipt, aliquoting is necessary for multiple uses. Avoid repeated freeze-thaw cycles. |
| Storage Buffer | Tris/PBS-based buffer, 6% Trehalose, pH 8.0 |
| Reconstitution | It is recommended that the vial be briefly centrifuged prior to opening. Reconstitute the protein in deionized sterile water to a concentration of 0.1-1.0 mg/mL. Adding 5-50% of glycerol (final concentration) and aliquot for long-term storage at -20℃/-80℃ is recommended. |
| Gene Name | G |
| Synonyms | G; Glycoprotein |
| UniProt ID | Q9QJT6 |
Product Specs
Note: While we prioritize shipping the format currently in stock, please specify your format preference in order remarks for customized preparation.
Note: All proteins are shipped with standard blue ice packs unless dry ice shipping is specifically requested and agreed upon in advance. Additional charges apply for dry ice shipping.
If you require a specific tag type, please inform us, and we will prioritize its development.
Target Background
KEGG: vg:1457772
Q&A
Basic Research Questions
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What experimental methods are used to express and purify recombinant SHRV-G?
Recombinant SHRV-G is typically expressed in E. coli cell-free systems or via reverse genetics in fish cell lines (e.g., EPC cells) . Key steps include:-
Cloning: Inserting the G gene (UniProt ID: Q9QJT6) into expression vectors under T7 or cytomegalovirus promoters .
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Purification: Using affinity chromatography (e.g., His-tag systems) followed by SDS-PAGE validation (>90% purity) .
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Functional verification: Binding assays (ELISA) to confirm receptor interaction .
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How is the structural integrity of recombinant SHRV-G validated?
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Fluorescent tagging: Coexpression with eGFP/mCherry in recombinant SHRV to track localization .
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Electron microscopy: Visualizing virion morphology and G protein spikes .
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Glycosylation analysis: Comparing migratory patterns in SDS-PAGE (noted absence in E. coli-expressed G due to prokaryotic systems) .
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Advanced Research Questions
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What challenges arise when designing chimeric viruses with heterologous G proteins?
Studies show that replacing SHRV-G with glycoproteins from related viruses (e.g., VHSV or IHNV) alters replication kinetics and host specificity . Key considerations:-
Temperature sensitivity: SHRV replicates optimally at 28–31°C, while chimeras may lose thermostability .
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Fusion activity: G protein truncations or mutations disrupt membrane fusion, requiring site-directed mutagenesis to map functional domains .
Example data:
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How do conflicting findings about the NV gene impact studies on SHRV-G function?
The nonvirion (NV) gene, a hallmark of Novirhabdoviruses, has unclear roles in G protein trafficking or virulence: -
What strategies optimize recombinant SHRV-G for vaccine development?
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Multivalent antigens: Coexpression of G with other viral proteins (e.g., IHNV N or VHSV P) via dual-gene recombinant SHRV .
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Adjuvant compatibility: Testing TLR agonists (e.g., poly I:C) to enhance anti-G antibody titers in fish models .
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Thermostability engineering: Introducing disulfide bonds in G to sustain immunogenicity at high temperatures .
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