Qiu Jian-Hua, Li Li, Wang Chun-Ren, Chen Jia, Chen Ai-Hua, Zhai Yan-Qing
College of Animal Science and Technology, Heilongjiang August-First Land Reclamation University, Daqing 163319, China.
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2008 Jun 30;26(3):183-6, 190.
To explore sequence differentiation of ITS and 28S rDNA-LSU of Orientobilharzia turkestanicum from bovine and caprine hosts.
Adult worms of O. turkestanicum from the naturally infected cattle, sheep, cashmere goat and goat were collected and identified morphologically as O. turkestanicum according to existing keys and descriptions. The genomic DNA was extracted from parasites of different hosts. The internal transcribed spacer (ITS, contains ITS-1, 5.8S nuclear ribosomal DNA, ITS-2) and 28S nuclear ribosomal DNA-LSU were amplified by PCR, sequenced and analyzed by Chromans and DNASTAR softwares, and the RNA secondary structure of 28S rDNA-LSU was analyzed by DNAMAN software.
ITS-1, 5.8S rDNA, ITS-2 and 28S rDNA-LSU of O. turkestanicum from bovine and caprine hosts were 384, 159, 331 and 1 304 bp, respectively. ITS-1 and 5.8S rDNA of O. turkestanicum from different definitive hosts were identical; ITS-2 of O. turkestanicum from cattle, sheep and cashmere goat were identical, with one nucleotide variation compared with that of goat; 28S rDNA-LSU of O. turkestanicum from sheep and cashmere goat were identical, with two nucleotides variation compared with that of cattle and goat. The RNA secondary structure of 28S rDNA LSU of O. turkestanicum from caprine hosts were identical or similar, but with large variation compared with that of cattle.
The rDNA sequence from different definitive hosts shows nucleotide variations to some extent and the RNA secondary structure of 28S rDNA-LSU from caprine hosts shows large variation in comparison to that of bovine.
探究来自牛和羊宿主的土耳其斯坦东毕吸虫ITS和28S rDNA-LSU的序列差异。
收集自然感染牛、绵羊、绒山羊和山羊体内的土耳其斯坦东毕吸虫成虫,依据现有检索表和描述进行形态学鉴定。从不同宿主的寄生虫中提取基因组DNA。通过PCR扩增内部转录间隔区(ITS,包含ITS-1、5.8S核糖体DNA、ITS-2)和28S核糖体DNA-LSU,利用Chromans和DNASTAR软件进行测序与分析,并通过DNAMAN软件分析28S rDNA-LSU的RNA二级结构。
来自牛和羊宿主的土耳其斯坦东毕吸虫的ITS-1、5.8S rDNA、ITS-2和28S rDNA-LSU分别为384、159、331和1304 bp。来自不同终末宿主的土耳其斯坦东毕吸虫的ITS-1和5.8S rDNA相同;来自牛、绵羊和绒山羊的土耳其斯坦东毕吸虫的ITS-2相同,与山羊的相比有1个核苷酸变异;来自绵羊和绒山羊的土耳其斯坦东毕吸虫的28S rDNA-LSU相同,与牛和山羊的相比有2个核苷酸变异。来自羊宿主的土耳其斯坦东毕吸虫的28S rDNA LSU的RNA二级结构相同或相似,但与牛的相比有较大差异。
来自不同终末宿主的rDNA序列在一定程度上存在核苷酸变异,并且与牛相比,羊宿主的28S rDNA-LSU的RNA二级结构有较大差异。