表达黄色荧光蛋白和乙胺嘧啶抗性基因的转基因柔嫩艾美耳球虫的发育和致病性分析  

Analysis for Pathogenicity and Development of Transgenic Eimeria tenella Expressing both Yellow Fluorescent Protein and TgDHFR-TS Genes

作　　者：闫文朝[1,2] 王天奇[1] 索勋[3] 韩利方[1] 丁轲[1] 董发明[1] 张龙现[2] 

机构地区：[1]河南科技大学动物科技学院,洛阳471003 [2]河南农业大学牧医工程学院,郑州450002 [3]中国农业大学动物医学院,北京100193

出　　处：《畜牧兽医学报》2009年第12期1794-1798,共5页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家“863”计划专题课题项目(2006AA02Z458);河南科技大学博士科研启动基金资助(09001350)

摘　　要：为了研究外源基因表达对转基因柔嫩艾美耳球虫(Eimeria tenella)生物学特性的影响,对转基因球虫(TE1)和BJ株(野生强毒)的发育和致病性进行比较分析。结果显示,相对于BJ株,转基因球虫TE1株繁殖力下降约4倍,低剂量感染时致病性下降明显,但是高剂量感染时依然保持较强的致病性。另外,在荧光显微镜下发现,TE1有滋养体、第一代裂殖体/裂殖子、第二代裂殖体/裂殖子和大小配子体等内生发育虫体。孢子化卵囊内4个孢子囊并非全部表达黄色荧光蛋白。这些结果说明黄色荧光蛋白和乙胺嘧啶抗性基因的表达在一定程度上降低了转基因柔嫩艾美耳球虫致病性和繁殖力。在柔嫩艾美耳球虫合子减数分裂过程中,存在同源或异源染色体重组现象。The pathogenicity and development of transgenic E.tenella strain TE1,which could express yellow fluorescent protein（YFP） and dihydrofolate reductase thymidylate synthase derived from Toxoplasma gondii（TgDHFR-TS）,in chickens were compared with that of the parental strain BJ to determine the effect of foreign gene on transgenic parasite.Results indicated that the fecundity of transgenic parasite（TE1） reduced at least 4 times than that of BJ strain.Low dosage of TE1 strain induced less pathogenic to chickens than BJ strain,but chickens inoculated with high dosage of TE1 oocysts displayed severe pathogenicity and certain mortality as well as BJ strain.In addition,trophozoites,the first generation and the second generation meronts and merozoites,microgamonts and macrogamonts of transgenic parasite TE1 were seen under flurescent microscope as that of BJ strain.More interestingly,not all four sporonts in the sporulating transgenic oocysts express YFP.These findings suggest that expressions of YFP and TgDHFR-TS genes reduced pathogenicity and reproductive potential of transgenic E.tenella to some extent.Recombination between homologous or non-homologous chromosomes appears during zygotic meiosis in E.tenella.

关 键 词：柔嫩艾美耳球虫 转染 繁殖力 致病性 活疫苗载体 

分 类 号：S852.723[农业科学—基础兽医学]