INFORMATIVE ENZYME/PROBE COMBINATIONS FOR THE MULTILOCUS DNA FINGERPRINTING OF MARSUPIALS

摘要

DNA fingerprinting has become an invaluable tool in the study of population genetics, paternity success, and individual identification; however, the species specificity of some methods has made the wide-range screening of many different species very time-consuming. In this study we describe the development and application of reliable and informative DNA fingerprinting techniques in a range of marsupial species using three different restriction enzyme and two oligonucleotide probe combinations. Six species from four marsupial families, the koala (Phascolarctidae), tammar wallaby (Macropodidae), southern hairy-nosed wombat (Vombatidae), kowari, and dusky and brown marsupial mice (Dasyuridae) were examined. Restriction enzymes HinfI AluI and HaeIII were used in combination with the digoxygenin (DIG)-labelled oligonucleotide probes (CAC)(5) and (GGAT)(4). The combinations of HinfI/(GGAT)(4), AluI/(CAC)(5) and AluI/(GGAT)(4) were the most informative, providing highly resolved bands, low background, and the lowest band sharing between individuals. The genetic diversity evident within the different species showed a clear relationship between the level of band sharing and population size. The greatest levels of band sharing were found in the kowaris (80%), which were part of a long-term captive colony originating from a few founders, and the lowest levels of band sharing were found in the marsupial mice (30-35%) and tammar wallaby (45%), which were caught from large outbred wild populations. [References: 28]