Comparative Studies of Lepidopteran Baculovirus-Specific Protein FP25K: Development of a Novel Bombyx mori Nucleopolyhedrovirus-Based Vector with a Modified fp25K Gene
Comparative Studies of Lepidopteran Baculovirus-Specific Protein FP25K: Development of a Novel Bombyx mori Nucleopolyhedrovirus-Based Vector with a Modified fp25K Gene
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DOI：
10.1128/jvi.00099-10
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发表时间：
2010-05-01
影响因子：
5.4
通讯作者：
Katsuma, Susumu
中科院分区：
文献类型：
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作者：
Nakanishi, Tadashi;Goto, Chie;Katsuma, Susumu
鳞翅目杆状病毒特异性蛋白FP25K在感染周期中发挥多种作用，包括闭塞体（OBs）和芽化病毒（BVs）的产生、口腔感染和死后宿主降解。为了探究鳞翅目杆状病毒中FP25K蛋白的共同功能和特异性功能，我们对I类和II类核多角体病毒（npv）和颗粒病毒（GV）的FP25K蛋白进行了比较分析。利用重组家蚕npv (BmNPV)，我们发现来自npv的fp25K能够消除在缺乏功能性fp25K的BmNPV突变体感染中观察到的所有表型缺陷，但来自GV的fp25K没有显示出恢复口腔感染性和死后宿主降解的能力。我们还观察到，将加利福尼亚Autographa californica multiple NPV (AcMNPV) fp25K引入到BmNPV基因组中，可以增强OB和BV的产生。根据这些结果，我们利用AcMNPV fp25K构建了一种新的bmnpv表达载体，并检测了其在BmN细胞和家蚕幼虫中的表达潜力。结果表明，AcMNPV fp25K的引入显著提高了外源基因产物在培养细胞中的表达，缩短了从幼虫血淋巴中获得分泌重组蛋白的时间。
Lepidopteran baculovirus-specific protein FP25K performs many roles during the infection cycle, including functions in the production of occlusion bodies (OBs) and budded viruses (BVs), oral infection, and postmortem host degradation. To explore the common and specific functions of FP25K proteins among lepidopteran baculoviruses, we performed comparative analyses of FP25K proteins from group I and group II nucleopolyhedroviruses (NPVs) and granulovirus (GV). Using recombinant Bombyx mori NPVs (BmNPVs), we showed that the FP25Ks from NPVs were able to eliminate all the phenotypic defects observed in an infection with a BmNPV mutant lacking functional fp25K but that FP25K from GV did not show abilities to recover oral infectivity and postmortem host degradation. We also observed that introduction of Autographa californica multiple NPV (AcMNPV) fp25K into the BmNPV genome enhanced OB and BV production. According to these results, we generated a novel BmNPV-based expression vector with AcMNPV fp25K and examined its potential in BmN cells and B. mori larvae. Our results showed that the introduction of AcMNPV fp25K significantly increases the expression of foreign gene products in cultured cells and shortens the time for obtaining the secreted recombinant proteins from larval hemolymph.