Li Q, Zhu J H, Xie X M, Liu Q L
Department of Parasitology, Tianjin Medical College and Institute of Molecular Biology, Nankai University.
Chin Med J (Engl). 1991 Jun;104(6):460-6.
High molecular weight DNA samples free of contaminating proteins and RNA obtained from one isolate (of Guangzhou origin) of Cysticercus tenuicollis and five isolates (of Tianjin, Harbin, Lanzhou, Shenyang and Zhengzhou origin) of Cysticercus cellulosae were subjected to thermal denaturation, restriction endonuclease digestion, Southern blotting and hybridization analysis. C. cellulosae DNA showed a melting temperature (Tm) of 82 degrees C corresponding to a 31% GC content whereas C. tenuicollis DNA melted at 85 degrees C suggesting 38.3% GC content. Visual inspection of ethidium bromide-stained gel showed differences not only between the DNAs of the two species of Taeniid cestodes, but also among the five isolates of C. cellulosae. Furthermore, we used two fragments (1.9kb and 5.5kb) of HindIII-derived restriction fragments of C. cellulosae DNA (Harbin origin) and pTS10 as probes to hybridize the DNAs of the Taeniid cestodes from six origins to detect inter- and intra-species genetic variation, the restriction fragment length polymorphisms (RFLPs) were identified.
从一株细颈囊尾蚴(广州株)和五株猪囊尾蚴(天津株、哈尔滨株、兰州株、沈阳株和郑州株)中获取了不含污染蛋白和RNA的高分子量DNA样本,并对其进行热变性、限制性内切酶消化、Southern印迹和杂交分析。猪囊尾蚴DNA的解链温度(Tm)为82℃,对应的GC含量为31%,而细颈囊尾蚴DNA在85℃解链,表明GC含量为38.3%。对溴化乙锭染色凝胶的目视检查显示,不仅两种带绦虫的DNA之间存在差异,而且五株猪囊尾蚴之间也存在差异。此外,我们使用猪囊尾蚴DNA(哈尔滨株)HindIII酶切片段的两个片段(1.9kb和5.5kb)和pTS10作为探针,与六个来源的带绦虫DNA杂交,以检测种间和种内遗传变异,确定了限制性片段长度多态性(RFLP)。