中文摘要：

克隆表达JEVE蛋白Ⅲ区基因,序列测定正确后进行融合表达、纯化。采用逆转录聚合酶链反应（RT-PCR）从JEVcDNA中扩增Ⅲ区基因,用限制性内切酶消化后插入到PMD-18T载体,序列测定正确后,再亚克隆到融合表达载体pET32a中。转化大肠杆菌BL21（DE3）,目的基因经IPTG诱导,由T7启动子调控表达了氨基端带6个连续组氨酸残基的JEVEⅢ区蛋白。在变性条件下对目的蛋白进行纯化,获得了融合6个组氨酸残基的JEVEⅢ区蛋白,蛋白纯度大于80%。证实构建了JEVEⅢ基因的重组表达载体,并获得了高纯度的融合表达蛋白,为以后的深入研究奠定了基础。

英文摘要：

To clone JEV E Ⅲ gene and purify fusion protein of JEV E Ⅲ, the target gene was amplified by reverse polymerase chain reaction （RT-PCR） from genome of JEV SA14 strain, and was inserted into cloning vector PMD-18T. After sequencing the target gene was subcloned into expression vector pET32a. The recombined plasmid pET32a-Ⅲ was transformed into the host strain E. coli BL21 （DE3）. After induced by IPTG, the strain controlled by T7 promoter expressed the fused JEV E Ⅲ protein with a hexahistidine tail in its N-terminal. The fusion protein mainly expressed in the inclusion bodies as showed by SDS-PAGE analysis. We purified the fusion protein by His-tag affinity chromatographv under denaturing condition, and the purity of fusion protein was higher than 80%. The conclusion is that construction of the recombinant expressing plasmid lays a basis for further study of the JEV E In protein.