The total RNA was extracted from sheep ovarian tissue by TRIzol method, and the coding region sequence of STMN1 gene was amplified by RT-PCR. The recombinant bait expression vector pGBKT7-STMN1 was constructed by homologous recombination technology, and the self-activation activity, cytotoxicity and expression of the bait protein in yeast two-hybrid system were determined. Screening of the interacting host proteins was performed from the yeast two-hybrid cDNA library of sheep ovarian tissue cells with the bait plasmid pGBKT7-STMN1. The results showed that the plasmid pGBKT7-STMN1 was successfully constructed in the study, and had no self-activation activity and cytotoxicity to yeast cells, and could be normally expressed in yeast cells. Totally, 12 clones interacting with STMN1 were screened out, and the sequencing analysis and point-to-point revealed the two interacting proteins, CREB and FOXM1, which may be involved in the regulation of cell proliferation, cycle and apoptosis.