Related Experiment Video
Updated: Aug 13, 2026
09:13
Using Reverse Genetics to Manipulate the NSs Gene of the Rift Valley Fever Virus MP-12 Strain to Improve Vaccine Safety and Efficacy
Published on: November 1, 2011
17.4K
Recombinant Protein Production and Purification of Rift Valley Fever Virus Nucleoprotein from Escherichia coli
Daniel Desmecht1, Hani Boshra2
1Faculty of Veterinary Medicine, Department of Morphology and Pathology, FARAH Research Center, University of Liège, Liège, Belgium.
Methods in Molecular Biology (Clifton, N.J.)
|December 13, 2024
Summary
This study details a protocol for producing Rift Valley fever virus nucleoprotein using bacterial expression systems. This method yields highly antigenic recombinant protein for diagnostic and immunological applications.
Area of Science:
- Virology
- Molecular Biology
- Immunology
Background:
- Recombinant viral protein production is crucial for studying viral immunity and developing diagnostic tools.
- Bacterial expression systems offer a scalable method for synthesizing and purifying viral proteins.
Purpose of the Study:
- To describe a protocol for the recombinant expression and purification of Rift Valley fever virus nucleoprotein (RVFV N).
- To provide a method for generating milligram quantities of RVFV N for downstream applications.
Main Methods:
- Cloning of RVFV nucleoprotein cDNA into a bacterial expression vector with a fusion protein and poly-histidine tag.
- Bacterial transformation, culture, lysis, purification, and dialysis to obtain the recombinant protein.
Main Results:
- Successful production of recombinant RVFV nucleoprotein.
- The protocol allows for upscaling to produce milligram quantities of purified protein.
Conclusions:
- The described protocol enables efficient production of recombinant RVFV nucleoprotein.
- The purified protein is suitable for immunological studies and diagnostic test development.
Related Concept Videos
01:27Production of Antibiotics
Penicillin, one of the earliest and most widely used antibiotics, is produced industrially by the filamentous fungus Penicillium chrysogenum. Large stirred-tank bioreactors ranging from tens to hundreds of thousands of liters maintain tightly controlled temperature, pH, and dissolved oxygen conditions to support fungal metabolism and maximize antibiotic yield. Penicillin is a secondary metabolite, synthesized primarily during the stationary growth phase, which requires a carefully managed...
01:30Production of Pharmaceuticals
Industrial insulin production uses genetically engineered E. coli expressing a proinsulin gene controlled by a tryptophan promoter and containing a methionine linker for later cleavage. The cells also carry ampicillin resistance for selective growth. Seed cultures are stored at −80 °C and production begins by thawing a small amount to inoculate starter cultures, which are progressively scaled to a 50,000-L bioreactor. In the bioreactor, E. coli grow in nutrient-rich media under sterile, tightly...
01:18Bacterial Gastroenteritis
Bacterial gastroenteritis, characterized by diarrhea, abdominal cramps, and vomiting, is often caused by ingestion of contaminated food or water and is frequently associated with pathogenic Escherichia coli strains. These microbes exploit two principal mechanisms to inflict disease.Shiga toxin–producing E. coli, also referred to as STEC—notably O157:H7—release Shiga toxins that target ribosomes, blocking protein synthesis. The B subunit of the toxin binds the host glycolipid receptor...