Abstract: The research aimed to clone the bovine inhibin α subunit gene and construct inhibin pET32a-mat peptide expression vector. The bovine ovaries were collected from slaughterhouses of Dachang County in Hebei Province. The total RNA was extracted and reverse transcribed into cDNA. A pair of primers was designed by Primer 5.0 according to the known bovine inhibin α-subunit sequence (Genbank No.NM174094.4). Inhibin α subunit mat peptide gene was amplified by RT- PCR. Finally, cDNA of this matured peptide was further amplified and cloned into the BamH Ⅰ and Hind Ⅲ sites of pET32a expressing vector to generate the recombinant inhibin pET32a-mat peptide plasmid. The results showed that the inhibin α subunit gene was successfully constructed. The authors preliminarily obtained the recombinant plasmid of inhibin and laid foundation for the further study of prokaryotic expression of recombinant proteins.