广西三地旋毛虫基因差异研究

发布时间：2018-11-18 07:19

【摘要】： 目的:分析广西三地(龙州、南丹、德保)旋毛虫分离株与河南旋毛虫分离株之间的基因差异以及与Genbank中旋毛虫种的基因差异,判断各分离株之间的亲缘关系;阐述旋毛虫自然感染龙州果子狸宿主的部分生物学特征,为探讨广西旋毛虫病爆发的原因提供科学依据。方法:对广西旋毛虫3个分离株和河南旋毛虫分离株基因组DNA进行提取,选取核糖体DNA(rDNA)的5S rDNA基因、线粒体DNA(mtDNA)的细胞色素氧化酶1(COX1)基因和线粒体DNA大亚基(mt-lsrDNA)基因作为分子标记,对目的基因进行PCR扩增,并采用双向测序法或T-A克隆测序法对PCR产物进行测序。应用Clustal X软件对旋毛虫基因序列进行排列比较,应用Dnastar软件包比较序列的同源性。然后采用Kimura双参数法计算遗传距离,采用邻接法(N-J法)、非加权组平均法(UPGMA)和最大简约法(MP)分别构建系统发生树,并且与Genbank已知的旋毛虫相应基因序列进行对照比较。同时对龙州果子狸旋毛虫分离株进行囊包形态学、毒力和保姆细胞形成时间进行实验观察。结果:龙州旋毛虫分离株、南丹旋毛虫分离株及德保旋毛虫分离株与基因库中T.spiralis 5S rDNA的相似度为98.3%～99.1%,遗传距离为0.014～0.028。龙州旋毛虫分离株、南丹旋毛虫分离株及河南分离株与基因库中T.spiralis COX1基因的相似度为98.8%～99.0%,遗传距离为0.003～0.005。龙州旋毛虫分离株、河南分离株与基因库中T.spiralis的mt-lsrDNA相似度分别为99.8%和99.5%,二者与基因库中T.spiralis的遗传距离则分别为0.003和0.005。四株分离株之间的相似度及遗传距离进行比较,龙州旋毛虫分离株、南丹旋毛虫分离株、德保旋毛虫分离株5S rDNA的相似度为97.6%～98.8%,遗传距离为0.014～0.028。龙州旋毛虫分离株和南丹旋毛虫分离株、河南分
[Abstract]:Objective: to analyze the genetic differences between Trichinella spiralis isolates and Henan Trichinella isolates in three regions of Guangxi (Longzhou, Nandan, Debao), and to determine the genetic relationship between Trichinella spiralis and Trichinella spiralis species in Genbank. This paper describes some biological characteristics of Trichinella spiralis naturally infecting the host of civets in Longzhou, and provides scientific basis for exploring the causes of outbreak of trichinellosis in Guangxi. Methods: genomic DNA was extracted from three isolates of Trichinella spiralis from Guangxi and Henan Province. The 5s rDNA gene of ribosomal DNA (rDNA) was selected. Cytochrome oxidase 1 (COX1) gene and mitochondrial DNA large subunit (mt-lsrDNA) gene of mitochondrial DNA (mtDNA) were used as molecular markers to amplify the target gene by PCR. The PCR products were sequenced by two-way sequencing or T-A clone sequencing. The sequence of Trichinella spiralis gene was compared by Clustal X software and the homology of the sequence was compared by Dnastar software package. Then the genetic distance is calculated by Kimura two-parameter method, and the phylogenetic tree is constructed by using the adjacency method (N-J method), the non-weighted group average method (UPGMA) and the maximum reduction method (MP), respectively. The results were compared with the corresponding gene sequences of Trichinella spiralis known by Genbank. At the same time, the cysts morphology, virulence and nanny cell formation time were observed. Results: the similarity between the isolates of Trichinella spiralis, Trichinella nandanensis and Trichinella pyrrolimus and T.spiralis 5s rDNA in gene bank was 98.3 and 99.1, and the genetic distance was 0.014 and 0.028. The similarity between the isolates of Trichinella spiralis, Trichinella Nandan and Henan isolates and T.spiralis COX1 gene in gene bank was 98.8 ~ 99.0 and the genetic distance was 0.003 ~ 0.005. The mt-lsrDNA similarity between the isolates of Trichinella spiralis in Longzhou and Henan Province was 99.8% and 99.55.The genetic distance between them and T.spiralis in gene bank was 0.003 and 0.005, respectively. The similarity and genetic distance among the four isolates were compared. The similarity of 5S rDNA and genetic distance of Trichinella spiralis, Trichinella nandanensis and Trichinella Dobo were 97.698. 8 and 0. 014 0. 028, respectively. Isolates of Trichinella spiralis and Trichinella Nandan in Longzhou, Henan Province
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R383

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