Abstract: To detect DNA variation of bovine lactoferrin gene, the sequence of 5^'﹣regulatory region has been studied by PCR﹣SSCP. Among the five sub-sequences of this region, Blf 5^'﹣1(227bp)，Blf 5^'﹣3(175bp) and Blf 5^'﹣5（293bp）were found to be polymorphic. After sequencing the Blf 5^'﹣1, G→A and T→G transitions were identified at the sites of -926 and -915 respectively. In the Blf 5^'﹣3 fragment, a G insertion at -478 was found. Transversions of C→A, G→C at –28 and +33 respectively were also detected in the Blf 5^'﹣5 region. The frequencies of mutated alleles were 0.101,0.112 and 0.237, respectively, in the three regions. By applying an analysis tool of TFSEARCH ver1.3, we studied the potential elements and protein factors which bind to the 5^'﹣region, and discovered the different regulatory elements of pre-mutation and post-mutation.