鸭粪中环孢子虫18S rDNA部分基因和ITS-1基因的克隆与分析

Discovery and Identification of Cyclospora-like Organism from Duck

应用PCR技术对在鸭粪中发现的疑似环孢子虫的18 S rDNA部分基因和ITS-1+基因进行了扩增,将扩增出的片段纯化后连接至pMD-18T载体上,选取阳性克隆进行序列测定,并利用NCBI在线BLAST程序和MEGA 4软件对测序结果进行了同源性比较和系统发育树构建。结果显示,测得的18 SrDNA序列与环孢子虫的相似性最高(98%),且在系统树中位于同一分支上;测得的ITS-1序列高度特异,在GenBank中未发现同源性序列,可以确定其为环孢子虫的一个新种,暂命名为鸭源环孢子虫。

The partial 18 S rDNA and ITS-1＋ gene of the Cyclospora-like organism from duck were amplified using PCR technique,and the positive amplicons were cloned into pMD-18T vector.The insert was successfully sequenced,then the resulting sequence was searched for homology with NCBI Blast program,and the phylogenetic tree with MEGA 4 software.The results revealed that the duck-associated Cyclospora-like organism was identified as a new Cyclospora sp.based on the 18S rDNA gene phylogenetic analysis and the specific ITS-1 gene.