Wu Z, Nakada T, Nagano I, Takahashi Y
Department of Parasitology, Gifu University School of Medicine, Tsukasa 40, Gifu, 500-8705, Japan.
Mol Cell Probes. 2000 Oct;14(5):291-7. doi: 10.1006/mcpr.2000.0316.
A method was developed for gene fingerprinting, combining the principles of restriction fragment length polymorphism and single-strand conformational polymorphism (PCR-RFLP-SSCP). Taking advantage of this method, we analysed the genotypes of 20 isolates from five species of Trichinella (Trichinella spiralis, Trichinella britovi, Trichinella nativa, Trichinella murrelli and Trichinella pseudospiralis) and two uncertain genotypes (Trichinella T6 and Trichinella T8). Target genes for the analysis included three kinds of random amplified polymorphic DNA (RAPD), the gene encoding 43 kDa excretory-secretory protein, the gene encoding mitochondrial cytochrome c -oxidase subunit I and the gene of 18 S rRNA. The genotype revealed by this method was in good concordance with the taxonomy of Trichinella (six species and two uncertain genotypes which is currently accepted based on morphology, isozyme pattern and reproductive isolation). More important, this method revealed intraspecies polymorphism among geographical isolates of T. spiralis and T. britovi.
开发了一种基因指纹识别方法，该方法结合了限制性片段长度多态性和单链构象多态性原理（PCR-RFLP-SSCP）。利用此方法，我们分析了来自旋毛虫属五个种（旋毛形线虫、布氏旋毛虫、本地旋毛虫、莫氏旋毛虫和伪旋毛虫）以及两种不确定基因型（旋毛虫T6和旋毛虫T8）的20个分离株的基因型。分析的靶基因包括三种随机扩增多态性DNA（RAPD）、编码43 kDa排泄分泌蛋白的基因、编码线粒体细胞色素c氧化酶亚基I的基因以及18 S rRNA基因。该方法揭示的基因型与旋毛虫的分类学结果高度一致（基于形态学、同工酶模式和生殖隔离目前认可的六个种和两种不确定基因型）。更重要的是，该方法揭示了旋毛形线虫和布氏旋毛虫地理分离株之间的种内多态性。
