摘要： 为了研究BmK IT提高AcMNPV抗虫活性的作用机制,将BmK IT基因插入到AcMNPV中形成重组病毒。采用重组单价病毒AcMNPV-BmK IT(IE1)、AcMNPV-BmK IT(P10)、AcMNPV-BmK IT(PH)和一种重组双价病毒AcMNPV-BmK IT(P10)-vcath (PH),通过四唑盐比色法(MTT)和蛋白质印迹法(Western Blot)分析了BmK IT在AcMNPV 的3个启动子调控下对Sf9细胞增殖和细胞凋亡的影响,结果显示,感染病毒36,48 h BmK IT在不同启动子调控下表达量从高到低依次为PH、P10、IE1。同时分析了AcMNPV介导的BmK IT与组织蛋白酶的协同表达对昆虫Sf9细胞增殖和调亡的机制,结果表明,AcMNPV-BmK IT(P10)-vcath(PH)处理组对Sf9细胞抑制率比AcMNPV-BmK IT(P10)处理组平均提高了14.5%,凋亡相关蛋白c-Myc、cleaved-Caspase3、Bax表达量增加,Bcl-2表达量减少。

关键词: AcMNPV, BmK IT, 组织蛋白酶, 协同作用

Abstract: In order to investigate the mechanism underlying the BmK IT mediated insecticidal activity of AcMNPV, an excitatory insect toxin, BmK IT, gene was inserted into the geneome of AcMNPV to construct a recombinant baculovirus.The scorpion insect neurotoxin BmK IT and vcath gene were constructed into the genome of AcMNPV using Bac-to-Bac system.Effects of BmK IT under these promoters of AcMNPV on Sf9 cells were analyzed and it revealed that the expression level of BmK IT was high under the regulation of PH promoter at 36, 48 h after the virus infected.Effects of AcMNPV mediated BmK IT and vcath synergism expression on the proliferation of Sf9 cells was also analyzed.Compared to AcMNPV-BmK IT (P10), the inhibition rate of AcMNPV-BmK IT (P10)-vcath(PH) increased 14.5% averagely, the expression level of c-Myc, cleaved-Caspase3 and Bax was raised and the expression of Bcl-2 was reduced.

Key words: AcMNPV, BmK IT, Vcath, Synergy

中图分类号: 

• Q78