Hepatitis C Virus (HCV) is a significant global health concern, affecting millions of people worldwide. The virus is classified under the genus Hepacivirus within the family Flaviviridae. One of the critical components of HCV is the nonstructural protein 5B (NS5B), which functions as an RNA-dependent RNA polymerase (RdRp). This enzyme is essential for the replication of the viral RNA genome. The segment of NS5B spanning amino acids 2634 to 2752 is particularly noteworthy due to its role in the virus’s replication machinery and its potential as a target for antiviral therapies .
The recombinant form of HCV NS5B (2634-2752 a.a) is typically expressed in Escherichia coli (E. coli) systems. The process involves cloning the gene segment encoding the desired amino acid sequence into an expression vector, which is then introduced into E. coli cells. The bacteria are cultured under conditions that induce the expression of the recombinant protein. Following expression, the protein is purified using conventional chromatography techniques to achieve a purity level exceeding 90%, making it suitable for various biochemical analyses .
The NS5B protein catalyzes the polymerization of ribonucleoside triphosphates (rNTPs) during viral RNA replication. This process is crucial for the synthesis of new viral RNA strands, which are necessary for the production of new viral particles. The enzyme’s activity can be inhibited by nucleoside and non-nucleoside inhibitors, which bind to the active site or allosteric sites on the protein, thereby blocking its function. These inhibitors are a focal point in the development of antiviral drugs aimed at treating HCV infections .