首页> 美国卫生研究院文献>Journal of Virology >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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Comparative Studies of Lepidopteran Baculovirus-Specific Protein FP25K: Development of a Novel Bombyx mori Nucleopolyhedrovirus-Based Vector with a Modified fp25K Gene

机译:鳞翅目杆状病毒特异蛋白FP25K的比较研究:新型的家蚕基于核多角体病毒的新型载体的修饰fp25K基因的开发。

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摘要

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.
机译:鳞翅目杆状病毒特异蛋白FP25K在感染周期中起许多作用,包括在产生闭塞体(OB)和发芽病毒(BV),口腔感染和死后宿主降解中起作用。为了探索鳞翅目杆状病毒中FP25K蛋白的共同和特定功能,我们对第一和第二组核多角体病毒(NPV)和颗粒病毒(GV)的FP25K蛋白进行了比较分析。使用重组家蚕NPV(BmNPV),我们显示NPV的FP25K能够消除在缺乏功能性fp25K的BmNPV突变体的感染中观察到的所有表型缺陷,但GV的FP25K没有显示出恢复口腔感染性和验尸的能力主机退化。我们还观察到,将B.NPV基因组中的加州Autographa californica多重NPV(AcMNPV)fp25K引入可增强OB和BV的产生。根据这些结果,我们用AcMNPV fp25K生成了一个新的基于BmNPV的表达载体,并研究了其在BmN细胞和家蚕幼虫中的潜力。我们的结果表明,AcMNPV fp25K的引入显着增加了培养细胞中外源基因产物的表达,并缩短了从幼虫血淋巴中获得分泌的重组蛋白的时间。

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