菲莱氏温扬球虫18S rDNA基因的扩增与克隆分析  

PCR Amplification,Cloning and Sequencing of Partial 18S rDNA from Wenyonella philiplevinei

作　　者：程家林[1] 李国清[1] 徐前明[1] 岳彩玲[1] 高振永[1] 朱海波[1] 刘霞[1] 

机构地区：[1]华南农业大学兽医学院,广州510642

出　　处：《中国畜牧兽医》2010年第6期67-70,共4页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金(30671577)

摘　　要：菲莱氏温扬球虫是鸭球虫病的重要病原之一,为了寻找种特异性的遗传标记,本研究采集广东省某鸭场的新鲜鸭粪,通过Sheather’s蔗糖漂浮法收集球虫卵囊,经形态学鉴定为菲莱氏温扬球虫;提取该球虫DNA样品,经过PCR扩增,首次获得了该虫的18S rDNA基因部分片段;对该基因进行了克隆和测序,比较了该虫株与其他原虫的亲缘关系。结果显示,对菲莱氏温扬球虫序列与艾美耳科其他球虫关系较近,系统进化树分析属于艾美耳科的另一分支。表明18S rDNA基因在鸭菲莱氏温扬球虫的分类鉴定上是一种有效的分子标记。Wenyonella philiplevinei is one of important pathogens of duck coccidiasis.To seek specific genetic marker of the protozoan,the fresh stool was obtained from a duck house in Guangdong,then coccidial oocysts were collected by Sheather＇s sucrose floatation procedure,and these oocysts were identified as W.philiplevinei according to morphology.The partial 18S rDNA was amplified by PCR,cloned and sequenced after the genomic DNA was extracted.The genetic relationship between the isolate and other protozoan was compared.The results showed that the sequence was closer to other coccidians of Eimeriidae.Phylogenetic analysis indicated that duck-derived W.philiplevinei isolate belonged to another branch of Eimeriidae.It is concluded that the 18S rDNA sequence provided an useful genetic marker for the differentiation of W.philiplevinei from other related protozoa.

关 键 词：菲莱氏温扬球虫 形态学 18S RDNA PCR 

分 类 号：S852.72[农业科学—基础兽医学]