中文摘要：

将实验室已有的pET32a—JcHSP17．5重组载体，经测序验证后，转入大肠杆菌BL21（DE3）plysS中，不同温度条件下，用不同浓度异丙基硫代-D-半乳糖苷（IPTG）诱导重组蛋白表达，优化表达条件；选择镍离子亲和层析柱（Ni—NTA）纯化JcHSP-17．5蛋白，SDS—PAGE电泳检测蛋白纯度；通过热激聚合反应来鉴定蛋白的分子伴侣活性．结果表明，17℃，200r／min，0．5mMIPTG诱导13h为JcHSP-17．5蛋白的最佳诱导表达条件，使用200mmol／L咪唑缓冲液纯化蛋白后，每500mL菌液可得到1．345mg的电泳纯蛋白，该蛋白在高温（45℃）条4#-下能够阻止革果酸脱氧醢（MDH）废生聚合反应．轰现出蠕强的夯平律侣活性．

英文摘要：

The pET32a-JcHSP17.5 recombinant vector verified by sequencing was transformed into E. coli BL21 （DE3）. The recombinant bacteria were induced with different concentrations of IPTG at dif ferent temperatures to express JcHSP-17.5 protein. JcHSP17.5 protein was purified through nickel ion affinity chromatography column （Ni-NTA）, and SDS-PAGE electrophoresis was run to test protein pu rity. The molecular chaperone activity was identified by heat shock agglutination. The results showed that the optimized expression condition was at 17 ℃, 200 r/min, and 0.5 mmol/L IPTG for 13 h. About 1. a45 mg electrophoresis pure protein in 200 mmol/L imidazole buffer was obtained from 500 mL bacte ria liquid culture. This protein can prevent malate dehydrogenase （MDH） agglutination at the condition of high temperature （45 ℃）, and showed relative strong molecular chaperone activity.