HCV NS5B (2634-2752 a.a)

Hepatitis C Virus NS5B (2634-2752 a.a) Recombinant
Cat. No.
BT17805
Source
Escherichia Coli.
Synonyms
Appearance
Sterile filtered colorless solution.
Purity
Greater than 90% as determined by SDS-PAGE.
Usage
THE BioTek's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.
Shipped with Ice Packs
In Stock

Description

Recombinant Hepatitis C Virus NS5B produced in E. coli is a single polypeptide chain containing 155 amino acids (aa 2634-2752) and having a molecular mass of 17kDa (NCBI Accession # NP_671491).
Recombinant HCV NS5B is fused to a 36 amino acid His-tag at N-terminus & purified by proprietary chromatographic techniques.

Product Specs

Introduction
The Hepatitis C Virus (HCV) is a small, enveloped virus with a single-stranded RNA genome. It belongs to the Flaviviridae family and measures approximately 50nm in diameter. HCV is characterized by a high replication rate, producing about a trillion viral particles daily in an infected person. The virus's RNA polymerase lacks proofreading ability, leading to a high mutation rate. This contributes to HCV's ability to evade the host's immune system. HCV is classified into six genotypes (1-6), with multiple subtypes within each genotype. The prevalence and distribution of these genotypes vary globally. Genotype identification is clinically significant as it influences the potential response to interferon-based therapy and the required treatment duration. Genotypes 1 and 4 demonstrate lower responsiveness to interferon-based treatment compared to genotypes 2, 3, 5, and 6.
Description
Recombinant Hepatitis C Virus NS5B (2634-2752 a.a) is a protein produced in E. coli. This single polypeptide chain consists of 155 amino acids and has a molecular weight of 17kDa. The protein sequence is derived from the NS5B protein of HCV (NCBI Accession # NP_671491). The recombinant HCV NS5B protein is engineered with a 36 amino acid His-tag at the N-terminus to facilitate purification. The protein is purified using proprietary chromatographic techniques.
Physical Appearance
Clear, colorless solution that has been sterilized by filtration.
Formulation
The HCV NS5B protein solution is provided at a concentration of 0.25mg/ml in a buffer containing 20mM Tris-HCl (pH 8.0), 0.15M NaCl, 10% glycerol, and 1mM DTT.
Stability
For short-term storage (up to 2-4 weeks), the HCV NS5B protein should be stored at 4°C. For long-term storage, it is recommended to store the protein at -20°C. To ensure stability during long-term storage, the addition of a carrier protein (0.1% HSA or BSA) is advisable. Avoid repeated freezing and thawing of the protein solution.
Purity
The purity of the HCV NS5B protein is greater than 90% as determined by SDS-PAGE analysis.
Source
Escherichia Coli.
Amino Acid Sequence
MRGSHHHHHH GMASMTGGQQ MGRDLYDDDD KDRWGSPMGF SYDTRCFDST VTESDIRTEE AIYQCCDLDP QARVAIKSLT ERLYVGGPLT NSRGENCGYR RCRASGVLTT SCGNTLTCYI KARAACRAAG LQDCTMLVCG DDLVVICESA GVQED.

Product Science Overview

Introduction

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 .

Preparation Methods

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 .

Chemical Reactions Analysis

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 .

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