Expression of the X protein of hepatitis B virus in insect cells using recombinant baculoviruses

Abstract

The baculovirus system was used to express the X protein of human hepatitis B virus (HBV). The X open reading frames (X ORFs) from cloned viral DNA of the HBV subtypes ayw and adr were introduced into the genome of Autographa californica nuclear polyhedrosis virus (AcNPV). The HBV-DNA of subtype adr derived from a hepatocellular carcinoma contains an X ORF and a 5' extended preX/X ORF, which were both used to construct X recombinant baculoviruses. Infection of Sf9 insect cells with these recombinant viruses yielded large amounts of the respective X proteins. They were identified by a set of mouse monoclonal antibodies directed against different epitopes of the ayw X protein using immunoblotting techniques. A subpopulation of the X protein expressed is modified, thus raising the molecular weight from the expected size of 17 kD to 21 kD. Indirect immunofluorescence and immunoelectron microscopy was performed to characterize the subcellular distribution of the X protein expressed in Sf9 cells. Data are presented that it accumulates as large globular structures within the cytoplasm and the nucleus of the infected cells.