目的构建天花粉蛋白(TCS)突变体基因,并进行表达及纯化。方法应用计算机预测TCS分子上可能的抗原决定簇,并进行定点突变。以栝楼基因组DNA为模板,经PCR扩增突变基因,与pRSET-A表达载体连接,转化大肠杆菌BL21(DE3),经IPTG诱导表达,并对表达产物进行Ni-NTA层析纯化。结果目的蛋白在大肠杆菌中获得高效可溶性表达,表达产物经纯化后,得到均一的TCS突变体蛋白。结论已成功构建了TCS突变体基因,并获得高效表达。 Objective To construct TCS mutant gene, express and purify it. Methods Computers were used to predict possible antigenic determinants on TCS molecules and perform site-directed mutagenesis. The genomic DNA was used as a template and the mutant gene was amplified by PCR. The gene was ligated with pRSET-A expression vector and transformed into E. coli BL21 (DE3). The expression was induced by IPTG and the expressed product was purified by Ni-NTA chromatography. Results The target protein was highly soluble and soluble in E. coli. After purification, the homogenous TCS mutant protein was obtained. Conclusion The TCS mutant gene has been successfully constructed and expressed efficiently.