›› 2002, Vol. 45 ›› Issue (6): 743-747.

• 研究论文 • 上一篇    下一篇

重组保幼激素酯酶的纯化和生物学效应

章东方   

  • 出版日期:2002-12-20 发布日期:2002-12-20

Purification and biological effects of a recombinant juvenile hormone esterase

ZHANG Dong-Fang   

  • Online:2002-12-20 Published:2002-12-20

摘要: 培养昆虫细胞生产重组昆虫保幼激素酯酶时细胞培养液的蛋白质浓度为153.2~188.0 μg/mL。批量处理纯化重组保幼激素酯酶时酶蛋白活力回收率33%,效果与梯度分离方法相当,但简便快速,可作为大量分离纯化的第一步。重组保幼激素酯酶对烟草天蛾Manduca sexta幼虫的生物学活性测定结果验证了重组保幼激素酯酶对烟草天蛾幼虫和自身天然酶有相似的生物学活性。

关键词: 重组保幼激素酯酶, 生物测定, 烟草天蛾, 离子交换层析

Abstract: The protein concentrations of cell supernatants infected with recombinant AcNPV (Autographa californica nuclear polyhedrosis virus )containing wild type or mutant juvenile hormone esterase were between 153.32-188.0 μg/mL. The enzyme activity recovery rate was about 33%. The stepwise purification of the enzyme had similar results as the gradient method, but was more convenient for handling large volumes of cell supernatants. The cuticle blackening te st showed that the recombinant juvenile hormone esterase from Heliothis virescens had biological effects on tobacco horn moth larvae(Manduca sexta).

Key words: recombinant juvenile hormone esterase, biological assay, Manduca sexta, ion exchange chromatography