Okamoto M, Ueda H, Hayashi M, Oku Y, Kurosawa T, Kamiya M
Institute of Experimental Animal Sciences, Osaka University Medical School, Suita-shi, Japan.
J Vet Med Sci. 1995 Apr;57(2):267-72. doi: 10.1292/jvms.57.267.
DNA from T. taeniaeformis digested with the restriction endonuclease was hybridized with digoxigenated oligonucleotide probe (CAC)5. Metacestode and adult showed same clear multibanding patterns, which were characteristic of multilocus DNA fingerprinting. The fingerprinting patterns were quite different from those of the rodent hosts. Genetic variations in 4 laboratory-reared isolates of T. taeniaeformis, including 3 isolates which have been reported to be indistinguishable by infectivity, morphology and protein composition of metacestode, were investigated using this technique. Each of the 4 isolates exhibited isolate-specific fingerprinting patterns and were easily distinguished from one another, thus it was considered that (CAC)5 was a highly resolvable and informative probe for cestodes. However, it was also indicated that (CAC)5 was so sensitive that applying fingerprinting with (CAC)5 to taxonomical or phylogenetic analysis was limited where habitat of the host was restricted to the small area. In comparison to fingerprinting with 32P-labeled (CAC)5, fingerprinting with digoxigenated (CAC)5 represented more and sharper bands. It was considered that a digoxigenated probe was more useful for genetic analysis of cestodes.
用限制性内切酶消化的绦虫DNA与地高辛标记的寡核苷酸探针(CAC)5进行杂交。中绦期幼虫和成虫呈现出相同的清晰多带模式,这是多位点DNA指纹图谱的特征。这些指纹图谱与啮齿动物宿主的指纹图谱有很大不同。利用该技术研究了4株实验室饲养的绦虫分离株的遗传变异,其中包括3株据报道在感染性、中绦期幼虫的形态和蛋白质组成方面无法区分的分离株。4株分离株中的每一株都表现出分离株特异性的指纹图谱,并且很容易相互区分,因此认为(CAC)5是一种对绦虫具有高分辨率和信息量的探针。然而,研究还表明(CAC)5过于敏感,以至于在宿主栖息地局限于小区域的情况下,将(CAC)5指纹图谱应用于分类学或系统发育分析受到限制。与用32P标记的(CAC)5进行指纹图谱相比,用地高辛标记的(CAC)5进行指纹图谱显示出更多更清晰的条带。认为地高辛标记的探针对于绦虫的遗传分析更有用。