重组ASP-1蛋白的表达与佐剂活性分析

Expression and adjuvanticity analysis of ASP-1 recombinant protein

目的:本研究采用基因工程技术表达ASP-1重组蛋白,并对其作剂活性作用进行分析。方法:采用密码子优化合成技术获得大肠杆菌偏性的ASP-1基因,将目的基因克隆至原核表达PQE30载体,转化表达宿主菌进行诱导表达。用纯化的重组表达的ASP-1蛋白作为佐剂,用模式抗原OVA和HIV肽抗原进行进行小鼠免疫试验,分析重组ASP-1蛋白的佐剂活性。结果:通过重叠PCR方法成功合成ASP-1基因,并在大肠杆菌中得到表达。用纯化的重组蛋白为佐剂与模式抗原一起免疫小鼠,同时与用铝佐剂组进行对照研究。试验结果表明,重组ASP-1蛋白可诱导模式抗原产生体液和细胞免疫反应,其佐剂活性高于铝佐剂。结论:重组表达的ASP-1蛋白具有良好的免疫佐剂活性。

Objective： To express the ASP - 1 protein in E. coli and analyze the adjuvanticity of the recombinant protein. Methods ： The code - optimized gene of ASP - 1 for E. coli expression was synthesized by assembly PCR and The purified recombinant protein mixed with antigens was used to immunize mice for analysis of the adjuvanticity. Results： Recombinant ASP - 1 protein was expressed successfully in E. coli. The purified and refolded protein was used to immunize mice. Result showed the recombinant ASP - 1 protein may induce both humoral and cell mediated immunity. Conclusion：The recombinant ASP - 1 protein is a potential adjuvant.