【作者】 王姝雅；

【导师】 李淑红；

【作者基本信息】 吉林大学 ， 病原生物学， 2006， 硕士

【摘要】 本研究首先从临床分离阴道毛滴虫,用肝浸汤进行纯培养,提取虫体总核酸,1%琼脂糖凝胶电泳,初步筛选出一株携带阴道毛滴虫病毒的虫株;然后根据国外已发表的阴道毛滴虫序列设计了一对覆盖全长的特异性引物,对阴道毛滴虫病毒基因组进行了全长cDNA克隆并测序,测得我国阴道毛滴虫病毒基因组全长为4649bp。该序列编码两个阅读框(ORF),二个阅读框有14nt的重复,在重复区有一个7聚体的滑动体(C CTT TTT)。ORF1(nt290-2326)包含678个氨基酸,编码分子量为74.9KDa的病毒衣壳蛋白;ORF2(nt2310-4581)包含756个氨基酸,编码分子量为85.8KDa的RNA依赖的RNA聚合酶(RDRP)蛋白。本研究的结果为深入研究我国阴道毛滴虫病毒奠定了良好基础。更多还原

【Abstract】 Trichomonas vaginalis, which parasitizes in urogenital system, is one kind of flagellates causing vaginal inflammation. At present, the characterization of T.vaginalis has not been understood completely and the discovery of Trichomonas vaginalis virus(TVV) played a pivotal role for T.vaginalis study in molecular biology. Several papers suggested that there were different kinds of dsRNA viruses in T.vaginalis. The T.vaginalis with virus was isolated from patients in Changchun and culture was carried out in vitro, the full-length cDNA of Trichomonas vaginalis virus was sequenced and constructed.T.vaginalis cultured in vitro T.vaginalis isolated from Changchun patients were cultured successfully in liver medium in vitro. The T.vaginalis strain with virus which had a 5.5kb extra band except genomic DNA was selected by 1% agarose gel electrophoresis analysis on total nucleic acids extracted from T. vaginalis. The T.vaginalis strain selected was cultured under axenic condition. The body configuration of paradises was observed by light and electron microscope. Growth state was very well in ox liver medium (pH=5.54) and the vital force didn’t decrease after 50 generation. The strain freezed in 10% DMSO was cultured continuously after recovery and the更多还原