柔嫩艾美耳球虫穿孔素样蛋白基因的原核表达与亚细胞定位  

Prokaryotic expression and subcellular localization of perforin-like protein gene for Eimeria tenella

作　　者：王文青 蒋保余 曲自刚[1] 刘保红[1] 肖星星[1] 马雪婷[1] 许笑 蔡建平[1] WANG Wen-qing;JIANG Bao-yu;QU Zi-gang;LIU Bao-hong;XIAO Xing-xing;MA Xue-ting;XU Xiao;CAI Jian-ping(State Key Laboratory of Veterinary Etiological Biology/Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China;College of Veterinary Medicine,Gansu Agricultural University,Lanzhou 730070,China)

机构地区：[1]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室,甘肃兰州730046 [2]甘肃农业大学动物医学院,甘肃兰州730070

出　　处：《中国兽医科学》2019年第11期1406-1414,共9页Chinese Veterinary Science

基　　金：国家重点研发计划项目（2017YFD0500403）

摘　　要：为了研究柔嫩艾美耳球虫(E.tenella)穿孔素样蛋白(EtPLP)在球虫感染中与宿主相互作用关系,首次成功获得EtPLP1(1932 bp)和EtPLP2(4215 bp)2个目的基因。通过表达纯化得到2个大小分别为37和67 ku的目的蛋白,Western-blot结果显示其反应原性良好。通过免疫荧光技术,观察到EtPLP1定位在子孢子顶端。mRNA转录水平检测结果显示,EtPLP1在子孢子阶段的表达量最高,其次是第1代裂殖子阶段,EtPLP2 mRNA水平在第1代裂殖子阶段表达量最高,其次是第2代裂殖子阶段;它们在其他阶段表达量极低或不表达。结果表明EtPLP1和EtPLP2可能参与虫体入侵与逸出宿主细胞,为EtPLPs蛋白功能研究奠定了基础。In order to study the interplay between perforin-like proteins for Eimeria tenella and the host cells during the infection,two target genes EtPLP1(1932 bp)and EtPLP2(4215 bp)were obtained.And then two corresponding proteins were detected with size of 37 ku and 67 ku respectively.And the Western-blot showed that both of them had good reactionogenicity.Using the immunofluorescence technique,EtPLP1 was localized to the apex of sporozoites.And the mRNA expression level was superior in the stage of sporozoite and then was the stage of the first generation merozoite.For EtPLP2,the expression level was superior in the stage of first generation merozoite,then was the stage of second generation merozoite.The expression levels were very low or none in the other stages for both EtPLP1 and EtPLP2.All the above results revealed that both EtPLP1 and EtPLP2 may involved in the invasion and escape for E.tenella to the host cells which established foundations to study the functions of EtPLPs.

关 键 词：柔嫩艾美耳球虫 穿孔素样蛋白 MRNA水平 亚细胞定位 

分 类 号：S852.723[农业科学—基础兽医学]