猪MAVS基因原核表达载体的构建及其多克隆抗体的制备
发布时间：2019-06-16 16:29
【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。
[Abstract]:The MAVS (mitochondrial antiviral signaling protein) gene was amplified and cloned from PK-15 cell line by RT-PCR, and the prokaryotic expression vector p ET-MAVS220, was constructed to transform the receptive cell Rosetta (DE3). The recombinant MAVS was induced by IPTG and immunized with recombinant MAVS for 4 weeks old Kunming mice to prepare polyclonal antibody against mitochondrial antiviral signal protein (MAVS). The best conditions for inducing expression were IPTG 0.05 mmol/L,37 鈩，
本文编号：2500667
[Abstract]:The MAVS (mitochondrial antiviral signaling protein) gene was amplified and cloned from PK-15 cell line by RT-PCR, and the prokaryotic expression vector p ET-MAVS220, was constructed to transform the receptive cell Rosetta (DE3). The recombinant MAVS was induced by IPTG and immunized with recombinant MAVS for 4 weeks old Kunming mice to prepare polyclonal antibody against mitochondrial antiviral signal protein (MAVS). The best conditions for inducing expression were IPTG 0.05 mmol/L,37 鈩，
本文编号：2500667
本文链接：https://www.wllwen.com/kejilunwen/jiyingongcheng/2500667.html
最近更新
教材专著
