斜纹夜蛾保幼激素环氧水解酶基因的克隆和原核表达

摘要/Abstract

摘要： 为分析斜纹夜蛾（Spodoptera litura）保幼激素环氧水解酶 (Juvenile hormone epoxide hydrolase,JHEH）基因在其生长发育过程中的功能，采用反转录聚合酶链式反应（RT-PCR）扩增SlJHEH基因的开放阅读框，并进行原核表达。SlJHEH基因开放阅读框为1 389 bp，编码462个氨基酸，预测蛋白质分子质量为52 ku，等电点为8.68。氨基酸序列存在催化三联体Asp226、Glu402和His429氨基酸残基及阴氧离子洞Tyr297、Tyr372和HGWP花样结构氨基酸残基。系统发育树分析结果表明，SlJHEH与棉铃虫（Helicoverpa armigera）JHEH亲缘关系最近，氨基酸序列同源性高达78%。将SlJHEH的编码区连接到原核表达载体pET32a上，成功构建了重组载体pET32a-SlJHEH,转入大肠杆菌BL21（DE3）感受态细胞中进行诱导表达。对表达产物进行SDS-PAGE和Western blotting检测，结果显示，斜纹夜蛾JHEH基因在大肠杆菌中得到高效且准确的表达。

关键词: 斜纹夜蛾, 保幼激素环氧水解酶, 克隆, 序列分析, 原核表达, 重组蛋白

张丽丽，武怡琼，杨正飞，郭丽，魏佳平，阚云超. 斜纹夜蛾保幼激素环氧水解酶基因的克隆和原核表达[J]. 河南农业科学, 2018, 47(12): 84-89.

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