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目的：用大肠杆菌融合蛋白表达载体pMAL—c2构建人血小板生成素突变体融合蛋白表达质粒pMAL-TPOM。方法：采用基因重组和表达、SD-聚丙烯酰胺凝胶电泳和蛋白纯化技术。结果：SDS-PAGE分析表明，IPTG诱导5h后大肠杆菌JM109表达到最高水平，约占大肠杆菌菌体总体蛋白的36．6％。除了大肠杆菌RR1不表达以外，大肠杆菌DH5a、H101均有较高水平的表达，表达产物血小板生成素突变体融合蛋白经SephacyIS-200和DEAE-SepharoseFF层析纯化后，蛋白纯度约为87．6％。结论：人血小板生成素突变体融合蛋白在大肠杆菌JM109、DH5a和H101中均获得高效表达。
OBJECTIVE: To construct the recombinant plasmid pMAL-TPOM of human thrombopoietin mutant by using E. coli fusion protein expression vector pMAL-c2. Methods: The gene recombination and expression, SD-polyacrylamide gel electrophoresis and protein purification techniques were used. Results: SDS-PAGE analysis showed that E. coli JM109 was expressed at the highest level 5 hours after induced by IPTG, accounting for 36.6% of the total E. coli bacteria protein. Escherichia coli DH5a and H101 were expressed at high levels except for the expression of Escherichia coli RR1, and the purity of the expressed product of the thrombopoietin mutant fusion protein was about 87.6 after purification by SephacyIS-200 and DEAE-Sepharose FF chromatography %. Conclusion: Human thrombopoietin mutant fusion protein was highly expressed in Escherichia coli JM109, DH5a and H101.