中文摘要：

通过RT-PCR从PK-15细胞系中扩增克隆MAVS（mitochondrial antiviral signaling protein）基因,构建原核表达载体p ET-MAVS220,转化感受态细胞Rosetta（DE3）,利用IPTG诱导表达,重组MAVS经纯化后免疫4周龄昆明系小鼠制备抗线粒体抗病病毒信号蛋白（MAVS）多克隆抗体。诱导表达的最佳条件为IPTG 0.05 mmol/L,37℃诱导6 h,重组MAVS以可溶性蛋白和包涵体两种形式表达。应用该重组蛋白免疫小鼠获得的抗MAVS多克隆抗体与纯化的重组MAVS蛋白反应效价可达1∶16 000;该抗体与Poly（I∶C）刺激PK-15细胞产生的MAVS及与转染了重组MAVS基因真核表达载体pcDNA3.0-MAVS的BHK-21细胞表达的MAVS蛋白发生特异性反应,效价可达1∶1 000,特异性良好。

英文摘要：

The MAVS gene was amplified from PK-15 cells by RT-PCR and was then subcloned into the prokaryotic expression vector to construct pET-MAVS220. The pET-MAVS220 was transfected into an E. coli Rosetta（DE3） to express the recombinant MAVS via IPTG induction. The recombinant MAVS was purified and used to immunize the four-week Kunming mice to prepare anti-MAVS polyclonal antibody. The constructed recombinant vector pET-MAVS220 was transformed into E. coli Rosetta（DE3） cells and the MAVS was expressed by induction of 0.05 mM IPTG at 37 ℃ for 6 h. The recombinant MAVS was identified in two forms of soluble protein and/or inclusion body. The purified MAVS was used to immunize the four-week Kunming mice to prepare a polyclonal anti- body. The reactivity of the anti-MAVS polyclonal antibody with prokaryotic expressed MAVS was identified to reach a high dilution of 1 ： 16000, and reach a dilution of 1：1000 with MAVS expressed in eukaryotic PK-15 cells by poly（I：C） and in BHK-21 cells transfected with the eukaryotic vector pcDNA3.0-MAVS. The anti-swine MAVS polyclonal antibody showed a high reactivity and specificity.