PCR amplification of a locus with RFLP alleles specific to African honey bees

An anonymous honey bee locus, detected previously with a cloned probe, has HhaI RFLP alleles specific to African bees or common to both African and European bees. To facilitate identification of these alleles, this region, 1231, was made analyzable with the PCR. The two halves of the region, excluding the termini, were amplified as two overlapping segments. Restriction sites were mapped, and the site differences responsible for the allelic RFLP patterns were determined. In the first half of the region, two polymorphic HhaI sites are present in the common alleles, whereas one, the other, or both of the sites are absent in the African alleles. In the second half, a third polymorphic HhaI site is present or absent in both common and African alleles. A short part of the second half of the region, including more of the terminus, was amplified as a third segment. Within this segment, close to this terminus, a fourth polymorphic HhaI site is absent in some African alleles.