The aim of this work is the expression of the PreS2-S region of surface antigen of hepatitis B virus (HBV) in yeast Pichia pastoris. A cDNA fragment encoding the Pres2-S protein of HBV was cloned to yeast transfer vectors. Based on cloned new plasmids pPIC3.5-PreS2-S (8707 bp) and pPIC9-PreS2-S (8980 bp) the recombinant strains of P. pastoris producing the PreS2-S region of surface antigen of HBV were obtained. The PAGE electrophoresis and immunoblotting of obtained recombinant PreS2-S protein confirm the molecular weight (34 kDa) and high specificity to the HBV antibodies.