Rapid communication: a PstI restriction fragment length polymorphism in the ovine Aconitase 2 gene.

Polymorphism. PstI restriction fragment length polymorphism (RFLP) in the ovine Aconitase 2 (AC02) gene. Source and Description of Probe. The probe is a .9kb AC02 cDNA from a porcine myocardial hgt 10 library (Zheng et al., 19901, cloned into the EcoRI site of pUC118. The insert was gel-purified and radiolabeled with [ c Y ~ ~ P I ~ C T P by the random primer method to specific activity > lo9 dpdpg. Hybridization Conditions. Nylon membranes were hybridized at 42°C in 5x SSC and 50% formamide according to established protocols (Ryan et al., 1993). Posthybridization washes were to a final stringency of . l x SSC at 60°C. Polymorphism. Genomic DNA from 26 feral Barbados x Rambouillet sheep (M.C. King and T. Rowell, Department of Integrative Biology, University of California-Berkeley) were screened with BamHI, BgZII, EcoRI, HindIII, MspI, PstI, and TaqI. Polymorphisms were only observed with PstI in this population. Frequency. Allelic frequencies for the 3.7and 3.2-kb PstI fragments (Figure 1 j in 26 animals were .45 and .55, respectively. Inheritance. The segregation of the PstI fragments was not followed because the pedigree structure of these feral animals was unknown. Chromosomal Location. Ovine Aconitase 2 has been mapped to bovine chromosome (BTA) 5 (Womack et al., 1993). Based on chromosome banding homology between cattle and sheep and the assignment of other BTA 5 loci to ovine chromosome (OAR) 3q (Burkin et al., 19931, the predicted chromosomal location of AC02 is OAR 3q. Probe Availability. Contact Howard Zalkin, Department of Biochemistry, Purdue University, West Lafayette, IN 47907. Comments. Aconitase, a member of the Krebs cycle system of enzymes catalyzing the interconversion of