In order to identify the role of CBF transcription factors in plant resistance of almond. The CBF1 homologue gene named AF-PdCBF1 (Genbank Accession No. KM245570) was isolated from the leaf of almond ‘Aifeng’(Prunus dulcis) in Xinjiang with the technology and method of PCR. Sequence analysis revealed that the open reading frame was 729 bp in full-length and encodes a protein of 242 amino acids, and its relative molecular mass was approximately 27.405 kDa, and AF-PdCBF1 has no signal peptide. The homology tree showed that AF-PdCBF1 with Prunus avium and Prunus mume has recently genetic relationship. The recombinant prokaryotic expression vector pET-PdCBF1 was constructed by inserting the DNA fragment into the prokaryotic expression vector pET-32a(+), and then transformed into E.coli Rosetta (DE3). The SDS-PAGE displayed that the expressed proteins consistented with the size of expected protein and then 47.8 kDa protein was obtained. The optimal expression condition was set 0.2 mmol/L IPTG,induced temperature 28℃, and induced time 4 hours. The results have provided a foundation for further purifying and identifying target protein and studying the function of AF-PdCBF1.