Production of a soluble and secreted antigenic fragment of HBsAg in yeast.
We have produced a fragment of hepatitis B surface antigen (HBsAg) corresponding to amino acids 1-60 as a fusion protein with the alpha mating factor of yeast. The product is secreted from yeast as a soluble monomer that expresses HBsAg antigenicity. Unlike other heterologous fusion proteins, it is not processed by the Lys-Arg endoprotease, possibly due to a proline in the linker between the two coding sequences. The resulting soluble fragment will enable us to map the immunodominant sites of HBsAg recognized by T cells and to identify additional factors contributing to vaccine potency.
Baba
E H
EH
Departamento de Bioquimica-ICB-UFMG, ICEB-Universidade Federal de Ouro Preto, Brazil.
Berkower
I
I
D000595:Amino Acid Sequence; D001483:Base Sequence; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D006514:Hepatitis B Surface Antigens / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D006515:Hepatitis B virus / Q000235:genetics; D000072235:Mating Factor; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis; D010452:Peptide Biosynthesis; D010446:Peptide Fragments / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D010455:Peptides / Q000235:genetics* / Q000302:isolation & purification; D010675:Pheromones / Q000235:genetics; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D014170:Transformation, Genetic
D004279:DNA, Viral; D006514:Hepatitis B Surface Antigens; D009838:Oligodeoxyribonucleotides; D010446:Peptide Fragments; D010455:Peptides; D010675:Pheromones; D011993:Recombinant Fusion Proteins; D000072235:Mating Factor