Objective To investigate the feasibility of PLCζ as an activator and its effect on fertilization and embryo development for oocytes after mouse round spermatid injection. Methods The recombinant plasmid pCRII-TOPOPLCζ was constructed in vitro and its expression was induced. The protein was identified by mass spectrometry and the antigenicity was detected by Western blot. Recombinant PLCζ protein was used as an activator of mouse ROSI oocytes, and fertilization and embryo development were recorded and analyzed statistically. Results The recombinant plasmid pCRIITOPO-PLCζ was successfully constructed, and the recombinant PLCζ protein was identified by mass spectrometry and had good antigenicity. The fertilization rate, two- to four-cell rate, eight-cell rate, and cystic rate associated with the recombinant PLCζ protein did not differ significantly from those in the blank group ( P > 0. 05). Conclusions The recombinant PLCζ protein has no significant effect on oocyte fertilization and embryo development in this study. The feasibility of being an oocyte activator after mouse ROSI is questionable.