中文摘要：

目的构建细粒棘球蚴（Echinococcus granulosus，EG）2热休克蛋白70（heat shock protein，2HSPT0）表达重组质粒，原核表达及纯化重组蛋白，并对重组蛋白进行免疫学特性的初步鉴定。方法从EG2HSP70／pGEM-T／JM109质粒中分离EG2HSP70目的基因，将此片段重组入表达载体pGEX-6P-1后转化入大肠杆菌BL21，鉴定后以IPTG诱导表达，通过SDS-PAGE检测其表达水平，用亲和层析纯化并分离重组蛋白，利用Western blot来鉴定重组蛋白的免疫学特性。结果酶切鉴定及测序分析表明，成功构建细粒棘球蚴pGEX-6P-1／EG2HSP70重组质粒；IPTG诱导表达后，融合蛋白占菌体总蛋白的39％，占裂解物上清总蛋白的70．4%，表明重组的EG2HSP70基因能在BL21中高效表达，表达的蛋白大部分以可溶性状态存在。亲和层析法纯化重组蛋白EG2HSP70，通过Western blot证实该重组蛋白能被细粒棘球蚴囊壁免疫兔血清识别。结论成功构建细粒棘球蚴中国大陆株pGEX-6P-1／EG2HSP70重组质粒，重组的EG2HSP70能够高效表达，表达的EG2HSP70具有免疫学活性。

英文摘要：

To construct a recombinant plasmid carrying heat shock protein 70 from Echinococcus granulosus, to express prokaryotically, to purify the recombinant protein and to identify the immunogenicity of its recombinant protein, the recombinant protein was obtained after the EGZHSP70 gene had been separated from EGZHSP70/ pGEM-T/JM109,inserted into expressive plasmid vector pGEX-6P-1 and expressed in E. coli BL21 under the induction of IPTG. Results of the induced expression level was detected by SDS-PAGE. The recombinant protein was purified by affinity chromatography and its biological activity was tested by Western blot analysis. The recombinant plasmid pGEX-6P-I/EG2HSP70 of Echinococcus granulosus has been constructed successfully and confirmed by restriction endonuclease analysis and DNA sequencing analysis. The expressed protein detected by SDS-PAGE was up to 39 % in total bacterical protein and 70.4 % in the total protein of cell lysate supernatants respectively,which demonstrated the recombinant EG2 HSP70 gene could be expressed in BL21 cells with high efficiency and the expressed proteins were mainly soluble. The recombinant protein was purified by affinity chromatography. Western blot revealed that the recombinant protein could be recognized by the sera from the rabbits immunized with EG. The recombinant plasmid EG2HSP70/pGEX-6P-1 of Echinococcus granulosus was successfully constructed and EG2HSP70 gene could be expressed in BL21 cells with high efficiency. Also the expressed proteins possess higher immunogenicity.