柔嫩艾美耳球虫组蛋白去乙酰化酶2A基因的克隆表达及转录动态分析  

Cloning and expression profiling of sirtuin-like histone deacetylase 2A during Eimeria tenella developmental cycle

作　　者：左云云[1,2] 马雪婷[1,2] 龚振兴[1,2] 殷昊[1,2] 刘保红[1,2] 韩政岚[1,2] 蔡建平[1,2] ZUO Yunyun MA Xueting GONG Zhenxing YIN Hao LIU Baohong HAN Zhenlan CAI Jianping(State Key Laboratory of Veterinary Etiological Biology/Key Laboratory of Veterinary Parasitology Gansu Province/Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yanzhou 225009, 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/甘肃省动物寄生虫病重点实验室,甘肃兰州730046 [2]江苏省动物重要疫病与人兽共患病防控协同创新中心,江苏扬州225009

出　　处：《畜牧与兽医》2016年第10期7-12,共6页Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金(31272554;31302084)

摘　　要：为研究柔嫩艾美耳球虫(Eimeria tenella)组蛋白去乙酰化酶2A(Et Sir2A)的生化功能及其对生活史发育的潜在调节作用,以柔嫩艾美耳球虫甘肃株为例,检测其在不同发育阶段表达量的差异。以基因组数据预测的编码序列为模板设计引物,采用RT-PCR的方法扩增得到Et Sir2A的全长ORF,构建p MAL-C2X-Et Sir2A重组表达质粒并进行原核表达;提取未孢子化卵囊、孢子化7h卵囊、孢子化卵囊、子孢子和第二代裂殖子5个发育阶段/时期的总RNA,采用qRT-PCR方法检测Et Sir2A的转录动态变化。测序结果显示,克隆的Et Sir2A ORF序列全长909 bp,编码302个氨基酸,并能在大肠杆菌原核系统中成功表达;qRT-PCR结果显示,Et Sir2A在不同发育阶段mRNA转录水平差异显著,在未孢子化卵囊阶段最高,第二代裂殖子阶段最低。结果为进一步研究Et Sir2A功能和开发新型抗球虫药物提供了数据基础。In order to characterize the biochemical features and potential epigenetic regulatory roles of sirtuin- like histone deacetylase 2A（ Et Sir2A） during Eimeria tenella developmental cycle,its gene cloning and transcriptional profiling were carried out using unsporulated,sporulating for 7 hours,sporulated oocysts,sporozoites,and second merozoite from E. tenella Gansu Strain. Based on the predicted ORF sequences from E. tenella,the primers for specific gene cloning were designed and qRT- PCR was performed. The cloned ORF sequence of Et Sir2 A is 909 bp which encodes 302 amino acids,and can be steadily expressed in Escherichia coli. The qRT- PCR results reveal that the mRNA transcription levels of Et Sir2 A were significantly in dynamic alterations during the main developmental stages of E. tenella,and the highest transcriptional level was in unsporulated oocysts. Our results revealed that Et Sir2 A could have potentially important regulatory roles during Eimerian life cycle,and will contribute to the novel anticoccidial development.

关 键 词：柔嫩艾美耳球虫 沉默信息调节因子2A 表达差异 QRT-PCR 

分 类 号：S855.9[农业科学—临床兽医学] Q936[农业科学—兽医学]