Hantaviruses are rodent-borne agents that cause severe human diseases. The coding sequences for the authentic and a His-tagged Puumala hantavirus (PUUV) nucleocapsid (N) protein were expressed in yeast (Saccharomyces cerevisiae). N-specific monoclonal antibodies demonstrated native antigenicity of the two proteins. All bank voles vaccinated with the His-tagged N protein in Freund's adjuvant (n=12) were defined as completely protected against subsequent virus challenge, based on the absence of viral N protein, RNA and G2-specific antibodies. In the group vaccinated with the yeast-expressed authentic N protein in Freund's adjuvant, 2/6 animals were defined as completely protected and 4/6 as partially protected. Moreover, when animals were vaccinated with the His-tagged N protein in an adjuvant certified for human use (alum), all (n=8) were at least partially protected (six completely, two partially). The general advantages of the yeast expression system make the described recombinant proteins promising candidate vaccines against hantavirus infection.