The polymorphism of the insulin-like growth factor binding protein 3 (IGFBP3) gene in Nanyang cattle, Luxi cattle and Chinese Simmental was analyzed by PCR-SSCP. The results showed that there were no polymorphism detected by the exon 3 of the bovine IGFBP3, and there were three genotypes (AA, AB and BB) detected by the exon 2 of the bovine IGFBP3 in three breeds. The frequency of A allele was  higher than that of B allele in three breeds. The frequency of AA genotype was the highest in three breeds, which Chinese Simmental was 0.6615, the Luxi cattle and Nangyang cattle were 0.4043 and 0.3095 respectively. The polymorphism fragments amplified by primer 1 (the exon 2 of the bovine IGFBP3) were cloned and sequenced. The sequencing results indicated that there was one single nucleotide mutation: T→C at 8 069 bp of IGFBP3 gene, and this mutation resulted in amino acid change: Phe→Leu.