Mitochondrial DNA control region sequences and seven microsatellites were used to estimate the genetic structuring, evolutionary history and historic migration patterns of the kob antelope (Kobus kob). Ten populations were analysed, representing the three recognized K. kob subspecies: K. k. kob in west Africa, K. k. thomasi in Uganda and K. k. leucotis in Sudan and Ethiopia. Despite being classified as K. k. thomasi and being phenotypically identical to the kob in Queen Elizabeth National Park (NP), the Murchison Falls population in Uganda showed high genetic similarity with the phenotypically distinct K. k. leucotis populations in Sudan and Ethiopia. This was regardless of marker type. Pairwise comparisons and genetic distances between populations grouped Murchison with K. k. leucotis, as did the Bayesian analysis, which failed to find any genetic structuring within the group. We propose that the divergent phenotype and life-history adaptations of K. k. leucotis reflect the isolation of kob populations in refugia in west and east Africa during the Pleistocene. Subsequent dispersal has led to secondary contact and hybridization in northern Uganda between lineages, which was supported by high levels of genetic diversity in Murchison. The reduced variability observed in Queen Elizabeth NP reflects a small founder population from west Africa and in part the decimation of Uganda's wildlife during the country's political turmoil in the 1970s. Due to similarities in phenotype and ecology, and the joint evolutionary history of their mtDNA sequences, the taxonomic status of K. k. kob and K. k. thomasi as separate subspecies is called into question.