Carnaby S, Butcher P D, Summerbell C D, Naeem A, Farthing M J
Digestive Diseases Research Centre, Medical College of St. Bartholomew's Hospital, London, UK.
Gene. 1995 Dec 1;166(1):167-72. doi: 10.1016/0378-1119(95)00572-5.
DNA fingerprinting has been widely used for genetic characterization and individual recognition in a range of species, from man and other mammals down the evolutionary scale to some lower eukaryotic parasites. These techniques utilise repetitive elements first characterised in the human genome, known as minisatellites, which display extensive allelic variability. Few biological or biochemical characteristics have been found that distinguish isolates of Giardia lamblia (Gl), or their apparent variations in virulence. We have characterized 21 Gl isolates in axenic culture using DNA fingerprinting with the human minisatellite probes, 33.6 and 33.15. Up to 12 variable bands per isolate were recognized in the size range of 2.5 to 15 kb by Southern blot hybridization of restriction endonuclease-digested Gl DNA. Most isolates demonstrated a distinct banding pattern or DNA fingerprint. The results suggest that this method may provide a basis for the detailed genotypic characterization of Gl which will be amenable to computer and statistical analysis for use in studies of virulence and epidemiology. Also, as Gl occupies a unique phylogenetic position as a member of the earliest known divergence from the eukaryotic line of descent, this study may provide a useful model for the study of other important eukaryotic pathogens, as it is rapidly becoming apparent that minisatellites are ubiquitous components of eukaryotic genomes.
DNA指纹技术已被广泛应用于一系列物种的遗传特征分析和个体识别,从人类和其他哺乳动物到进化尺度较低的一些低等真核寄生虫。这些技术利用了最早在人类基因组中发现的重复元件,即微卫星,它们表现出广泛的等位基因变异性。很少有生物学或生化特征能够区分蓝氏贾第鞭毛虫(Gl)的分离株,或者它们在毒力方面的明显差异。我们使用人类微卫星探针33.6和33.15,通过DNA指纹技术对21株处于无菌培养状态的Gl分离株进行了特征分析。通过对限制性内切酶消化后的Gl DNA进行Southern印迹杂交,在2.5至15 kb的大小范围内,每个分离株最多可识别出12条可变带。大多数分离株表现出独特的条带模式或DNA指纹。结果表明,该方法可能为Gl的详细基因型特征分析提供基础,这将便于进行计算机和统计分析,用于毒力和流行病学研究。此外,由于Gl作为已知最早从真核生物谱系分化出来的成员之一,占据着独特的系统发育位置,这项研究可能为其他重要的真核病原体研究提供一个有用的模型,因为越来越明显的是,微卫星是真核基因组中普遍存在的组成部分。