SpltNPV和SeNPV几丁质酶基因大肠杆菌表达产物对病毒的增效作用

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为明确杆状病毒几丁质酶对病毒杀虫剂的增效作用,采用PCR扩增获得斜纹夜蛾核型多角体病毒(SpltNPV)不含N-端信号肽序列的几丁质酶基因片段SpltNPVChiA/S-和甜菜夜蛾核型多角体病毒(SeNPV)几丁质酶基因编码区全序列SeNPVChiA,分别克隆入原核表达载体pET-28a,转化大肠杆菌BL21(DE3),获得高效表达,表达蛋白的分子量为62 kD和66 kD.将含有表达产物SphNPVChiA/S-和SeNPVChiA的大肠杆菌菌液分别与SphNPV或SeNPV一起加入人工饲料,饲喂3龄斜纹夜蛾或甜菜夜蛾幼虫,两种幼虫取食后致死时间均显著缩短,其中,SpltNPV+Spit-NPVChiA/S-和SphNPV+SeNPVChiA的LT50比SphNPV对照处理缩短0.90天和0.89天,LT90缩短1.90天和1.97天;SeNPV+SphNPVChiA/S-和SeNPV+SeNPVChiA的LT50比SeNPV对照处理缩短0.47天和0.50天,LT90缩短0.57天和0.59天;而pET-28a/BL21对病毒的侵染无明显增效作用.两种病毒几丁质酶基因的大肠杆菌表达产物对病毒早期侵染均有明显的增效作用,但同源几丁质酶和异源几丁质酶对病毒的增效作用没有显著差异.
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