Expression and characterization of the multiplied, recombinant preS1 antigen of hepatitis B virus.
The amino acid sequence encoded by the preS1 region of hepatitis B virus genome is expressed on the surface of virions and subviral particles. The preS1 region is involved in the recognition of specific receptors responsible for the attachment of HBV to the host cell. The cell receptor binding site was assigned to the preS1 (20-47 aa) fragment. In order to obtain a large quantity of preS1 binding domains of HBV the expression vector pWX4 was constructed. It contains four tandemly joined DNA sequences, each coding for preS1 (20-49 aa), fused with the 3' end of a DNA fragment coding for 450 aa of beta-galactosidase. E. coli cells transformed with this vector produce fusion protein beta-gal-preSlx4 in the form of inclusion bodies. Owing to the specific trypsin digestion, the preSlx4 domain was cleaved from the fusion protein. The resulting product, a 16 kDa protein, was isolated and purified by anion exchange chromatography. The presence of four Asp-Pro bonds in this sequence and the primary structure of the first 28 N-terminal amino acids were determined. Following the confirmation of the antigenic properties, the recombinant preS1 protein was used for detection of the anti-preS1 response in sera from HBV infected patients.
Sidorkiewicz
M
M
Department of Biochemistry, Medical University of Lódz, Poland.
Płucienniczak
G
G
Płucienniczak
A
A
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D006509:Hepatitis B / Q000276:immunology; D006510:Hepatitis B Antibodies / Q000276:immunology; D006514:Hepatitis B Surface Antigens / Q000235:genetics* / Q000276:immunology; D006801:Humans; D008969:Molecular Sequence Data; D011498:Protein Precursors / Q000235:genetics* / Q000276:immunology; D011993:Recombinant Fusion Proteins; D012091:Repetitive Sequences, Nucleic Acid; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology
D006510:Hepatitis B Antibodies; D006514:Hepatitis B Surface Antigens; D011498:Protein Precursors; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; C050806:presurface protein 1, hepatitis B surface antigen