Fowler E V, Houlden B A, Sherwin W B, Hoeben P, Timms P
Centre for Molecular Biotechnology, School of Life Science, Queensland University of Technology, Brisbane, Australia.
Biochem Genet. 1998 Jun;36(5-6):193-206. doi: 10.1023/a:1018720707514.
Highly repeatable randomly amplified polymorphic DNA (RAPD) markers were developed for parentage studies in the koala (Phascolarctos cinereus). Of the 25 RAPD primers screened, 5 (20.0%) produced 32 repeatable polymorphic RAPD bands (average/primer = 6.4 +/- 4.2). A high level of polymorphism was observed for each group of koalas (Featherdale, 71.9%; Lone Pine, 84.4%). All 25 koalas could be uniquely identified using either RAPD or microsatellite markers. Of the 32 RAPD markers generated in koalas, 25 were informative for parentage analyses. These RAPD markers successfully determined both parents to three offspring and a male parent to a fourth offspring. Paternity analysis (where the female parent is known) succeeded in assigning the correct male parent to seven offspring. Our RAPD-PCR method generates informative genetic markers that are useful for parentage determination and individual identification of captive koalas. This would provide genetic analysis to zoos and wildlife parks as a low-cost alternative to the more expensive microsatellite markers.
为了对考拉（树袋熊，学名：Phascolarctos cinereus）进行亲子关系研究，开发了高度可重复的随机扩增多态性DNA（RAPD）标记。在筛选的25个RAPD引物中，5个（20.0%）产生了32条可重复的多态性RAPD条带（平均每个引物 = 6.4 ± 4.2）。对每组考拉（费瑟戴尔动物园的考拉，多态性水平为71.9%；孤松动物园的考拉，多态性水平为84.4%）都观察到了高度的多态性。使用RAPD或微卫星标记可以唯一地识别所有25只考拉。在考拉中产生的32个RAPD标记中，有25个对亲子关系分析具有信息价值。这些RAPD标记成功地确定了三只后代的双亲以及第四只后代的雄性亲本。亲权分析（已知雌性亲本）成功地为七只后代确定了正确的雄性亲本。我们的RAPD-PCR方法产生了可用于圈养考拉亲子关系确定和个体识别的信息性遗传标记。这将为动物园和野生动物园提供遗传分析，作为比更昂贵的微卫星标记成本更低的替代方法。
