Resumo

RUBIO, Ivonne; COMBITA, Alba Lucía; ORTIZ-REYES, Blanca  e  NAVAS, María-Cristina. Hepatitis C virus Core protein production and purification in baculovirus expresión system for biological assays. Biomédica [online]. 2005, vol.25, n.1, pp.34-45. ISSN 0120-4157.

 Introduction: The hepatitis C virus (HCV) commonly causes persistent infection. One of the viral mechanisms that could be involved in avoiding viral clearance is the ability of HCV to induce functional alterations of the immune system cells, specifically of dendritic cells (DCs). The studies to identify the viral proteins involved in DCs functional alterations have attributed these effects to the HCV Core protein, the capsid structural unit. Objective: One of the limitations to evaluate Core protein properties is the unavailability to obtain and purify the complete protein (191 aa). The aim of this study was to produce and purify the recombinant Core protein in a eukaryotic system, to evaluate the effect of the protein in human DCs cultures. Results: The Core protein p23 isoform was expressed in a baculovirus system, and then purified using isoelectric point separation and electroelution. The purity of Core protein was confirmed by silver stain and western blot. These analyses showed the presence of two bands that correspond to p23 and p21 isoforms of Core protein as previously reported. Conclusions: The protein obtained has several conditions of naïve Core protein as molecular weight, isoforms and subcellular localization. The procedures described in this paper could be applied to membrane associated proteins produced in eukaryotic systems.

Palavras-chave : Hepatitis C Virus; Core protein; Baculovirus; expression system; isoelectrofocusing; purification.

        · resumo em Espanhol     · texto em Espanhol     · Espanhol ( pdf )