乙型脑炎病毒NS1蛋白的可溶性表达与抗原性分析

Soluble expression and antigenic analysis of NS1 protein of Japanese encephalitis virus

为获得可溶性表达的乙型脑炎病毒(JEV)NS1蛋白,将编码JEV SA14-14-2株NS1蛋白基因克隆至原核表达载体pMAL-c5X中,构建重组融合表达质粒pc5X-NS1;用重组质粒转化宿主菌ER2523后,经IPTG诱导得到可溶性的融合蛋白MBP-NS1;优化诱导表达条件,于28℃、0.3 mmol/L IPTG诱导4 h;最后融合蛋白经直链淀粉树脂柱纯化。结果表明:重组融合蛋白MBP-NS1具有良好的抗原性和特异性。

To obtain the expression of NS1 protein of Japanese encephalitis virus＇in a soluble form, the NS1 gene of JEV strain SA14 - 14 -2 was cloned into the expression vector pMAL - c5X, then the recombinant plasmid pc5X - NS1 was confirmed by sequencing. After the recombi- nant plasmid was transformed into host bacteria ER2523, the fusion protein MBP - NS1 was expressed in the soluble form and purified by induced by isopropyl beta - D - thiogalactopyranoside （IPTG） , then the expression conditions were further optimized. Western - blot and indirect ELISA analysis showed that the recombinant fusion protein MBP - NS1 could be specific reactive with antibodies against JEV. This work can provide an important basis for further study on NS1 protein of Japanese encephalitis virus.