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| Home -> Products & Services -> Custom Plasmid Vector Construction Services -> Custom Construction Service of RABV Expression Plasmid Vector | ||
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Custom Construction Service of RABV Expression Plasmid Vector Rabies virus (RABV) belongs to the family Rhabdoviridae and is a single-stranded, negative-sense RNA virus. It is the causative agent of rabies, a disease primarily transmitted through the saliva of infected animals. Upon infection, the virus targets the central nervous system, leading to severe neurological symptoms and death. Although rabies is rare in humans, it remains a significant public health concern worldwide. The recombinant rabies virus (RV or RABV) enables scientists to enhance the exploration of neural circuitry and the progression of vaccine research by exploiting the virus's distinctive infection pathway.RGBiotech offers custom construction service for RABV expression plasmid vector. Through our recombinant rabies virus expression vector construction service, you can obtain high-quality and high-efficiency viral products to advance your research. Contact us at admin@rgbiotech.com to learn more about our services and customization options. Why Choose RGBiotech? 1) Vector Backbone: We provide two options for RABV vector backbones which are based on CVS-N2c strain and HEP-Flury strain, respectively.2) Professional Team: Our scientists have an in-depth understanding of theoretical knowledge in the field of recombinant RABV and are proficient in technical operations. They have extensive experience in the field of RABV expression vector construction and can efficiently solve various technical problems encountered in the project. 3) Quality Assurance: The constructed recombinant rabies virus expression vector is strictly verified by sequencing to ensure the accuracy of the sequence and guarantee the reliability of the product quality. 4) High-quality Products: Upon additional request, we can provide endotoxin-free plasmids suitable for the transfection of packaging cells, reducing the impact on cell transfection and virus packaging and improving the success rate of experiments. 5) Detailed Report: Along with the delivery of the product, a detailed report is provided, including COA and sequencing verification data, which provides a comprehensive reference for subsequent research. Read More Rabies Virus (RABV) belongs to the genus Lyssavirus of the family Rhabdoviridae and is the pathogen causing rabies. Rabies is an ancient and highly lethal zoonotic infectious disease. Once the disease occurs, the fatality rate is almost 100%. Every year, a large number of people around the world are at risk of infection due to being bitten by animals carrying the virus. It is mainly transmitted through saliva, posing a serious threat to public health security.Genomic Structure of RABV The RABV genome is a single-stranded negative-sense RNA, approximately 12 kb in length. From the 3' to the 5' end, it encodes five major proteins in sequence: nucleoprotein (N), phosphoprotein (P), matrix protein (M), glycoprotein (G), and RNA-dependent RNA polymerase (L). The N protein tightly wraps the RNA, forming a ribonucleoprotein complex (RNP), which protects the nucleic acid and participates in the initiation of viral transcription and replication. The P protein assists the L protein in its function and is involved in viral RNA synthesis. The M protein connects the viral nucleocapsid to the envelope and plays a key role in virus assembly and budding. The G protein is the only glycoprotein on the virus surface, mediating the binding of the virus to host cell receptors and membrane fusion, and determining the neurotropism and immunogenicity of the virus. The L protein has RNA polymerase, methyltransferase, and polyadenylation enzyme activities and is responsible for the transcription and replication of the viral genome. These proteins play critical roles in viral replication, transcription, and assembly.Applications of Recombinant Rabies Virus (RABV) Recombinant rabies virus has broad applications in scientific research and biotechnology, including but not limited to the following aspects.1) Neural Circuit Tracing: RABV can spread retrogradely across synapses and is used in neuroscience research to trace the connections between neurons and analyze the structure and function of complex neural circuits. 2) Vaccine Development: Construct recombinant viral vectors expressing the protective antigens of RABV for the development of efficient and safe new rabies vaccines, enhancing the immune effect and reducing the potential risks of traditional vaccines. 3) Gene Therapy Vector: Utilize the neurotropism of RABV, transform it into a gene therapy vector, and deliver therapeutic genes to the diseased sites of nervous system-related diseases (such as neurodegenerative diseases). Features of Recombinant Rabies Virus (RABV) 1) Strong Neurotropism & High Infection Efficiency: It can specifically infect nerve system cells, having a targeted advantage in nervous system research and the treatment of related diseases.2) Good Immunogenicity: Its antigens can effectively stimulate the body's immune response, making it suitable for vaccine development. 3) Capacity for Foreign Genes: Its genome can be modified through genetic engineering techniques, and foreign genes can be inserted to achieve specific functions. 4) Safety: Genetic engineering can reduce the virus's pathogenicity, enhancing experimental safety. Packaging Process of Recombinant Rabies Virus (RABV) 1) Vector Construction: Clone the target gene into the RABV expression vector to ensure the correct insertion of the foreign gene and the integrity of the expression elements.2)Helper Plasmid Preparation: Prepare helper plasmids expressing proteins required for RABV replication and packaging. Helper vectors typically contain genes encoding proteins essential for RABV replication and packaging (N, P, M, G, L). These proteins are indispensable during the viral packaging process. 3) Transfection of Packaging Cells: Use endotoxin-free plasmids suitable for the transfection of packaging cells to co-transfect the recombinant expression vector and the helper vector into specific packaging cells (such as BHK-21 cells). The helper vector provides other proteins required for virus packaging and helps assemble the recombinant virus. 4) Virus Cultivation and Amplification: After transfection, the cells are cultured under appropriate conditions. The virus replicates, assembles, and is released into the culture medium in the cells. The virus titer is amplified through multiple passages. 5) Virus Harvesting and Purification: Collect the culture medium containing the virus and purify the virus using methods such as ultracentrifugation and chromatography to remove impurities and unassembled viral components. 6) Titer Determination: Measure viral titer using fluorescent markers or qPCR. 7) Quality Control: Perform sequencing, Western blot, and other quality control assays to ensure viral quality and correct expression of the foreign gene. Packaging Limitation of Recombinant Rabies Virus (RABV) Generally speaking, the RABV expression vector can accommodate a certain length of foreign fragments. However, as the length of the foreign gene increases, the packaging efficiency and stability of the virus will be affected. Therefore, the recombinant RABV vector has size limitations for foreign gene insertion, typically ranging from 1.5 to 2.5 kb according to current experience. Insertions larger than this may affect viral packaging efficiency and infectivity.Elements of the Recombinant Rabies Virus (RABV) Expression Vector 1) Promoter: Such as the CMV promoter, which has strong transcriptional activity and drives the efficient expression of foreign genes in host cells.2) Multiple Cloning Site: It contains recognition sites for multiple restriction endonucleases, facilitating the insertion of foreign genes. 3) Terminator: Ensures that transcription terminates at the appropriate position to prevent the generation of abnormal transcripts. 4) Selection Marker: Such as antibiotic resistance genes (such as the ampicillin resistance gene, etc.), which is convenient for screening cell clones containing the recombinant vector. 5) RABV genome sequence: The N, P, M, G, and L genes from the RABV genome of CVS-N2c strain or HEP-Flury strain. 6) Helper Vector: The helper vector mainly encodes the essential proteins for the replication and packaging of RABV, such as the genes of N, P, L and other proteins. These proteins can interact synergistically with the recombinant expression vector in the packaging cells to make up for the insufficient expression of certain proteins of the recombinant virus due to the insertion of foreign genes, ensuring the normal assembly and release of the virus. |
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