黄艾美耳球虫纯种分离及ITS-1序列测定

摘要

为建立一种有效鉴定兔球虫的分子生物学方法,采用单卵囊分离技术,分离纯化黄艾美耳球虫.根据GenBank中发表的艾美耳属球虫18S rDNA和5.8S rDNA序列,设计特异性引物,建立PCR方法并针对黄艾美耳球虫第一内转录间隔区进行扩增,PCR产物直接测序.结果分离出黄艾美耳球虫,并扩增出包含黄艾美耳球虫第一内转录间隔区的基因条带,随后测定了该条带序列,大小为455bp.研究结果填补了兔球虫第一内转录间隔区序列遗传资料的空白,为兔球虫虫种及虫株的准确鉴定奠定了基础.%To establish a molecular biological method for identifying coccidium species,a pair of specific primers was designed according to previously published sequences of 18S rDNA and 5.8S rDNA of Eimeria coccidia in the GenBank database. The pure species of Eimeria flavescens was isolated using single-oocyst isolating technique and the gene of ITS - 1 of E. flavescens was amplified by PCR assay. The PCR product was cloned and sequenced,which showed that the pure species of E. flavescens was successfully isolated and the gene fragment including ITS - I of E. flavescens was 455 bp long.