目的 :构建重组人淋巴毒素α缺失体 (rhLT αΔN2 7)的原核表达载体 ,在大肠杆菌中进行表达 ,建立纯化rhLT αΔN2 7的工艺。方法 :从Jurkat细胞中提取总RNA ,用RT PCR扩增rhLT αΔN2 7基因 ,并插入原核表达载体 pET 2 3b中 ,转化大肠杆菌BL2 1(DE3) ,用IPTG诱导rhLT αΔN2 7表达。包涵体经洗涤和复性后 ,用DEAESepharoseFF和Phenyl SepharoseFF纯化。结果 :rhLT αΔN2 7以包涵体的形式表达 ,表达量占菌体总蛋白的 30 %以上。纯化后 ,rhLT αΔN2 7的纯度达 99% ,比活性高于 8× 10 7U/mg。纯化样品的相对分子质量 (Mr)、等电点 ,以及N端序列等其他理化性质均同预计的结果相符。结论 :构建了rhLT αΔN2 7的表达载体 ,并成功地在大肠杆菌中进行了表达 ,建立了rhLT αΔN2 7的纯化工艺 Objective: To construct prokaryotic expression vector of recombinant human lymphotoxin α (rhLT αΔN2 7) and express in E. coli, and to establish a method for purification of rhLT αΔN2 7. Methods: The total RNA was extracted from Jurkat cells. The rhLT αΔN2 7 gene was amplified by RT PCR and inserted into prokaryotic expression vector pET 2 3b. The recombinant plasmid was transformed into E. coli BL21 (DE3) and induced by IPTG. Inclusion bodies were washed and refolded and purified with DEA ​​Sepharose FF and Phenyl Sepharose FF. Results: rhLT αΔN2 7 was expressed in the form of inclusion bodies, and the expression level accounted for more than 30% of total bacterial proteins. After purification, rhLT αΔN2 7 purity of 99%, specific activity higher than 8 × 10 7U / mg. Purified samples relative molecular mass (Mr), isoelectric point, and N-terminal sequence and other physical and chemical properties are in line with the expected results. Conclusion: The expression vector of rhLT αΔN2 7 was constructed and successfully expressed in E. coli. The purification technology of rhLT αΔN2 7 was established