鸡柔嫩艾美耳球虫SO7基因的原核表达及动物免疫试验  

Prokaryotic and Expression of SO7 Gene from Chicken Eimeria tenella and Animal Immunization Test of Its Recombinant Protein

作　　者：仇保丰[1,2] 宋鸿雁[1,3] 严若峰[1] 宋小凯[1] 徐立新[1] 李祥瑞[1] 

机构地区：[1]南京农业大学动物医学院,江苏南京210095 [2]南通出入境检验检疫局,江苏南通226004 [3]南通大学实验动物中心,江苏南通226001

出　　处：《动物医学进展》2016年第2期1-5,共5页Progress In Veterinary Medicine

基　　金：国家自然科学基金项目(31372428;31201896)

摘　　要：为了研究鸡柔嫩艾美耳球虫SO7基因原核表达重组蛋白的免疫效力,分析其在鸡抗柔嫩艾美耳球虫感染中对鸡平均增重、抗球虫指数(ACI)、减少盲肠病变和卵囊数量中的作用,从柔嫩艾美耳球虫孢子化卵囊中提取总RNA作为模板,用RT-PCR扩增出SO7基因片段,再应用DNA重组技术将SO7基因克隆到原核表达载体pET32a(+)中并用IPTG诱导表达。SDS-PAGE结果显示,pET32a(+)-SO7表达的目的蛋白约为40ku,主要以包涵体形式存在。用纯化的重组蛋白进行动物免疫试验显示,SO7重组蛋白在鸡抗柔嫩艾美耳球虫感染中有较好的免疫保护作用,能够显著提高鸡柔嫩艾美耳球虫感染鸡的平均增重和抗球虫指数(ACI),对减少盲肠病变和卵囊数量也有部分作用。To study the immune efficacy of recombinant protein, which was obtained with the expression SO7 gene of Eimeria tenella in Escherichia coli, and evaluate the immunization effects of recombinant protein on the average body-weight gain, anti-coccididal index （ACI）, caecal lesions scores and oocyst out- put during the course of chickens against E.tenella infection. The SO7 gene was amplified from total RNA of E.tenella sporulated oocysts by RT-PCR. By the means of DNA recombination technique, SO7 gene was cloned into prokaryotic expression vector of pET32a（＋）, and the proteins were expressed by induc- tion of IPTG. As the results of SDS-PAGE indicated, pET32a（＋）-SO7 could express the SO7 gene, and most of the recombinant proteins were existed in the inclusion bodies with a molecular mass of approxi- mately 40 ku. Using the purified proteins, the animal immunization test was carried out, and the results showed that the recombinant protein of SO7 gene share an ideal immunoprotective effect, the average body-weight gain and the anticoccididal index of chickens challenged with Eimeria tenella could be signifi- cantly increased, and the caecal lesion scores and oocyst output could also be partly alleviated.

关 键 词：柔嫩艾美耳球虫 SO7基因 克隆 表达 免疫效力 

分 类 号：S852.723[农业科学—基础兽医学] Q786[农业科学—兽医学]